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18 pages, 5943 KiB  
Article
Rabies Realities: Navigating Barriers to Rabies Control in Rural Zambia—A Case Study of Manyinga and Mwansabombwe Districts
by Muma Chipo Misapa, Eugene C. Bwalya, Ladslav Moonga, Josiah Zimba, Emmanuel S. Kabwali, Mwenya Silombe, Edgar Chilanzi Mulwanda, Christopher Mulenga, Martin C. Simuunza, Hirofumi Sawa, Bernard Hang’ombe and Walter Muleya
Trop. Med. Infect. Dis. 2024, 9(7), 161; https://doi.org/10.3390/tropicalmed9070161 (registering DOI) - 18 Jul 2024
Abstract
Rabies persists as a longstanding issue in Zambia, despite being preventable. The current control measures, including dog vaccination, population control, and movement restriction, guided by ‘The Control of Dogs Act Chapter 247 of the Laws of Zambia’, have not yielded the desired impact [...] Read more.
Rabies persists as a longstanding issue in Zambia, despite being preventable. The current control measures, including dog vaccination, population control, and movement restriction, guided by ‘The Control of Dogs Act Chapter 247 of the Laws of Zambia’, have not yielded the desired impact in many areas of the country including Manyinga and Mwansabombwe districts. These two districts continue to report low dog vaccination rates, unrestricted dog movements, and escalating cases of animal and human rabies, along with dog bites. Aligned with global aspirations to achieve zero human rabies cases by 2030, this study scrutinizes the determinants and obstacles hampering the execution of rabies control initiatives in Manyinga and Mwansabombwe. Spanning approximately 11 months, this cross-sectional study gathered pre- and post-vaccination data from 301 households in Manyinga and 100 households in Mwansabombwe. Questionnaires probed knowledge, attitudes, and practices related to rabies prevention and control. A transect survey, key informant interviews, and assessment of rabies vaccination and dog bite records complemented the data collection. Findings revealed that 88.0% of respondents from both districts possessed knowledge about rabies, confirming affected species and transmission. Moreover, 76.8% in Manyinga and 88.6% in Mwansabombwe were acquainted with rabies prevention and control methods. Concerning dog owners, 89.0% were aware of rabies, 66.0% understood its prevention and control, and the majority identified bites as the primary mode of transmission. Despite the high level of knowledge recorded during the survey, the implementation of preventive measures was low, which was attributed to low levels of law enforcement by the local government authority, inadequate staffing in the veterinary department, unwillingness to pay for dog vaccinations, and unavailability of rabies vaccine at the veterinary office in both districts. Vaccination coverage stood at 64.0% in Manyinga and 21.0% in Mwansabombwe. Notably, education and occupation exhibited a positive significant association with rabies knowledge. In terms of dog bite cases, Manyinga recorded 538 dog bite cases from 2017 to June 2022, while Mwansabombwe recorded 81 dog bite and 23 jackal bite cases from 2021 to June 2022. The study underscores critical knowledge gaps in rural areas and emphasizes the imperative for enhanced public education and awareness programs, improved rabies surveillance, free mass vaccination campaigns, and community engagement to augment vaccination coverage and knowledge about rabies. Full article
(This article belongs to the Section Neglected and Emerging Tropical Diseases)
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<p>Manyinga district study area.</p>
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<p>Mwansabombwe district study area.</p>
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<p>Manyinga dog bite records from January 2017–30th June 2022.</p>
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<p>Mwansabombwe dog and jackal bites record from January 2021–30th June 2022.</p>
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12 pages, 933 KiB  
Article
Resource Utilization of Residual Organic Sludge Generated from Bioenergy Facilities Using Hermetia illucens Larvae
by Kyu-Shik Lee, Eun-Young Yun and Tae-Won Goo
Insects 2024, 15(7), 541; https://doi.org/10.3390/insects15070541 (registering DOI) - 18 Jul 2024
Abstract
Residual organic sludge generated from bioenergy facilities (BF-rOS) is often disposed instead of recycled, thus contributing to further environmental pollution. This study explored the resource utilization of BF-rOS using Hermetia illucens larvae (BSFL). When BF-rOS was fed to BSFL for two weeks, the [...] Read more.
Residual organic sludge generated from bioenergy facilities (BF-rOS) is often disposed instead of recycled, thus contributing to further environmental pollution. This study explored the resource utilization of BF-rOS using Hermetia illucens larvae (BSFL). When BF-rOS was fed to BSFL for two weeks, the dry weight per individual BSFL was approximately 15% of that of BSFL that were fed food waste (FW). However, the dry weight increased by approximately two-fold in BSFL that were fed effective microorganism (EM)-supplemented BF-rOS containing 60% moisture. However, under both conditions, the BSFL did not mature into pupae. In contrast, the highest dry weight per BSFL was observed with the BF-rOS/FW (50%:50%) mixture, regardless of EM supplementation. Furthermore, the highest bioconversion rate was observed when the BSFL were fed the BF-rOS/FW (50%:50%) mixture, and the frass produced by the BSFL contained fertilizer-appropriate components. In addition, the nutritional components of the BSFL exhibited a nutrient profile suitable for animal feed, except for those fed BF-rOS only. In conclusion, this investigation demonstrates that BF-rOS should be recycled for fertilizer production by mixing it with FW as a BSFL feed, which generates the valuable insect biomass as potential nutrition for animal feeding. Full article
(This article belongs to the Special Issue Insects and Their Derivatives for Human Practical Uses 2nd Edition)
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<p>Effect of moisture in residual organic sludge generated from bioenergy facilities (BF-rOS) and effective microorganism (EM) on the survival and body weight of <span class="html-italic">Hermetia illucens</span> (black soldier fly larvae) (BSFL). (<b>A</b>) Six-day-old BSFL were inoculated into BF-rOS with varying moisture levels and allowed to grow for 14 days. Food waste (FW) served as a positive control. The statistical significance of each experimental group was determined by comparing it to the control. # indicates <span class="html-italic">p</span> &lt; 0.0001. (<b>B</b>) Six-day-old BSFL were inoculated into BF-rOS with varying moisture and EM levels and allowed to grow for 14 days. The statistical significance of each experimental group was determined by comparing it to the control. * indicates <span class="html-italic">p</span> &lt; 0.05. (<b>A</b>,<b>B</b>) Surviving BSFL were harvested and counted to determine survival rate. They were dried to measure the dry weight/larva. Dry weight/larva was calculated by dividing total dry weight of BSFL by the total number of BSFL. All experiments were conducted independently in triplicate. Values are presented as the mean ± standard deviation of survival rate or dry weight/larva.</p>
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<p>Effect of BF-rOS/FW mixtures and EM on the survival and body weight of BSFL. BSFL were grown for 14 days after inoculating 6-day-old BSFL, and then we collected and counted the number of surviving BSFL to determine survival rate. They were dried to measure the dry weight/larva. Dry weight/larva was determined by dividing total dry weight of BSFL by the total number of BSFL. All experiments were conducted independently in triplicate. Values are presented as the mean ± standard deviation of survival rate or dry weight/larva. The statistical significance of each experimental group was determined by comparing it to FW alone. * and *** indicate <span class="html-italic">p</span> &lt; 0.05 and <span class="html-italic">p</span> &lt; 0.001, respectively.</p>
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<p>Effect of BSFL powder on animal cell survival. (<b>A</b>) Survival of human intestinal cell line Caco-2. (<b>B</b>) Survival of mouse fibroblast cell line L929. (<b>A</b>,<b>B</b>) Cell survival of BSFL powder-treated cells was determined as a percentage of the survival of untreated control cells. No significant differences were observed in any of the samples. OS indicates BF-rOS.</p>
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14 pages, 2588 KiB  
Article
Genome-Wide Association Studies of Growth Trait Heterosis in Crossbred Meat Rabbits
by Zhanjun Xiao, Yuchao Li, Li Yang, Mingyan Cui, Zicheng Wang, Wenqiang Sun, Jie Wang, Shiyi Chen, Songjia Lai and Xianbo Jia
Animals 2024, 14(14), 2096; https://doi.org/10.3390/ani14142096 (registering DOI) - 18 Jul 2024
Abstract
The application of heterosis can not only effectively improve the disease resistance and meat quality of livestock, but also significantly enhance the reproduction and growth of livestock and poultry. We conducted genome-wide association studies using data from F2 crossbred meat rabbits to screen [...] Read more.
The application of heterosis can not only effectively improve the disease resistance and meat quality of livestock, but also significantly enhance the reproduction and growth of livestock and poultry. We conducted genome-wide association studies using data from F2 crossbred meat rabbits to screen out candidate genes with significant dominant effects associated with economic trait variation. High-throughput sequencing technology was used to obtain SNPs covering the whole genome to evaluate the homozygosity of the population genome, and analyze the number, length, frequency, and distribution of ROHs in the population. Candidate genes related to economic traits of meat rabbits were searched based on high-frequency ROH regions. After quality control filtering of genotype data, 380 F2 crossbred rabbits were identified with 78,579 SNPs and 42,018 ROHs on the autosomes. The fitting of the Logistic growth curve model showed that 49-day-old rabbits were a growth inflection point. Then, through genome-wide association studies, 10 SNP loci and seven growth trait candidate genes were found to be significantly related to body weight in meat rabbits at 84 days of age. In addition, we revealed the functional roles and locations of 20 candidate genes in the high-frequency ROH region associated with economic traits in meat rabbits. This study identified potential genes associated with growth and development in the high-frequency ROH region of meat rabbits. In this study, the identified candidate genes can be used as molecular markers for assisted selection in meat rabbits. At the same time, the inbreeding situation based on ROH assessment can provide reference for breeding and breeding preservation of meat rabbits. Full article
(This article belongs to the Section Animal Genetics and Genomics)
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<p>The weight frequency distribution of 380 samples of F2 generation rabbits at seven growth stages (35, 42, 49, 56, 63, 70, and 84 days old). Note: The histogram represents the frequency distribution, and the curve represents the normal distribution curve.</p>
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<p>(<b>A</b>) The number of SNPs on each autosome before and after quality control. (<b>B</b>) Autosome SNP density distribution plot. (<b>C</b>) Principal component analysis of SNPs.</p>
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<p>84-day old GWAS Manhattan chart and Q-Q chart. The X-axis in (<b>A</b>) is color-coded to represent different chromosomes, and the Y-axis represents the −log10(<span class="html-italic">p</span>) of the SNPs. The horizontal red line parallel to the <span class="html-italic">X</span>-axis represents the genome-wide significance level threshold. (<b>B</b>) Sites on the red line indicate genome-wide significance and are associated with target traits.</p>
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<p>(<b>A</b>) Scatterplot of total length of ROH per individual within the population. (<b>B</b>) Histogram of the number of short, medium, and long fragments of ROH. (<b>C</b>) Density distribution of ROHs. The shorter ROH segments (0.3~2 Mb) account for a high proportion. (<b>D</b>) Histogram of ROH distribution for different chromosomes.</p>
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<p>(<b>A</b>) Twenty-eight Gene Ontology entries (BP: biological process; CC: cellular component; MF: molecular function). (<b>B</b>) Eight important KEGG pathways.</p>
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14 pages, 820 KiB  
Article
Use of Different O/W or W/O Emulsions as Functional Ingredients to Reduce Fat Content and Improve Lipid Profile in Spanish Cured Processed Meat Product (‘Chorizo’)
by Elena Martínez, Wagner Goncalves Vieira Júnior, Manuel Álvarez-Ortí, Adrián Rabadán and José Emilio Pardo
Foods 2024, 13(14), 2262; https://doi.org/10.3390/foods13142262 (registering DOI) - 18 Jul 2024
Abstract
In this work, three out of five types of oil-in-water and water-in-oil emulsions were selected to replace pork backfat to reduce the fat content and the improve monounsaturated and polyunsaturated fatty acid content in dry cured sausage (‘chorizo’). Different characteristics of the new [...] Read more.
In this work, three out of five types of oil-in-water and water-in-oil emulsions were selected to replace pork backfat to reduce the fat content and the improve monounsaturated and polyunsaturated fatty acid content in dry cured sausage (‘chorizo’). Different characteristics of the new product were studied: the texture profile, color, nutritional value, lipid profile, vitamin E and thiobarbituric acid (TBA) and sensory qualities. The use of emulsions to replace the animal fat affected all color parameters, obtaining darker, less red and yellow products, which could impact the consumer’s purchase intention. This replacement also altered the texture parameters, increasing or decreasing the hardness in comparison with the control sample. The cohesiveness, however, decreased in all cases, which meant that when the samples are cut for consumption, they disintegrated more than the traditional ones. The most relevant aspect for nutritional value is that the use of the new emulsions helped to reduce the total fat and energy value of the reformulated samples. The most significant aspect is that this reformulation helped to reduce the fat content—specifically, saturated fat—while increasing the content of Omega 3/6. The new formulas contained significant quantities of TBA and vitamin E when comparing them with the traditional product. Full article
(This article belongs to the Section Meat)
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<p>‘Chorizo’ formulas used in the present work.</p>
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<p>Results obtained for the sensory analysis (external appearance, texture, smell and taste) of the different cured chorizo reformulations evaluated. E2CH: chia oil emulsion with guar gum and inulin; E2PM: pumpkin oil emulsion with guar gum and inulin; E3CH: chia oil emulsion with defatted chia flour and inulin; E3PM: pumpkin oil emulsion with defatted chia flour and inulin; E5CH: chia oil emulsion with HPMC and xanthan gum; E5PM: pumpkin oil emulsion with HPMC and xanthan gum.</p>
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20 pages, 5640 KiB  
Article
Whole-Genome Resequencing Analysis of the Camelus bactrianus (Bactrian Camel) Genome Identifies Mutations and Genes Affecting Milk Production Traits
by Huaibing Yao, Zhangyuan Pan, Wanpeng Ma, Zhongkai Zhao, Zhanqiang Su and Jie Yang
Int. J. Mol. Sci. 2024, 25(14), 7836; https://doi.org/10.3390/ijms25147836 (registering DOI) - 17 Jul 2024
Abstract
Milk production is an important trait that influences the economic value of camels. However, the genetic regulatory mechanisms underlying milk production in camels have not yet been elucidated. We aimed to identify candidate molecular markers that affect camel milk production. We classified Junggar [...] Read more.
Milk production is an important trait that influences the economic value of camels. However, the genetic regulatory mechanisms underlying milk production in camels have not yet been elucidated. We aimed to identify candidate molecular markers that affect camel milk production. We classified Junggar Bactrian camels (9–10-year-old) as low-yield (<1.96 kg/d) or high-yield (>2.75 kg/d) based on milk production performance. Milk fat (5.16 ± 0.51 g/100 g) and milk protein (3.59 ± 0.22 g/100 g) concentrations were significantly lower in high-yielding camels than those in low-yielding camels (6.21 ± 0.59 g/100 g, and 3.93 ± 0.27 g/100 g, respectively) (p < 0.01). There were no apparent differences in gland tissue morphology between the low- and high-production groups. Whole-genome resequencing of 12 low- and 12 high-yield camels was performed. The results of selection mapping methods, performed using two methods (FST and θπ), showed that 264 single nucleotide polymorphism sites (SNPs) overlapped between the two methods, identifying 181 genes. These genes were mainly associated with the regulation of oxytocin, estrogen, ErbB, Wnt, mTOR, PI3K-Akt, growth hormone synthesis/secretion/action, and MAPK signaling pathways. A total of 123 SNPs were selected, based on significantly associated genomic regions and important pathways for SNP genotyping, for verification in 521 additional Bactrian camels. This analysis showed that 13 SNPs were significantly associated with camel milk production yield and 18 SNPs were significantly associated with camel milk composition percentages. Most of these SNPs were located in coding regions of the genome. However, five and two important mutation sites were found in the introns of CSN2 (β-casein) and CSN3 (κ-casein), respectively. Among the candidate genes, NR4A1, ADCY8, PPARG, CSN2, and CSN3 have previously been well studied in dairy livestock. These observations provide a basis for understanding the molecular mechanisms underlying milk production in camels as well as genetic markers for breeding programs aimed at improving milk production. Full article
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<p>Differential physiological and biochemical indexes in high- and low-yielding camels. ** <span class="html-italic">p</span> &lt; 0.01; *** <span class="html-italic">p</span> &lt; 0.001; NS, not significant.</p>
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<p>Comparison of microscopic morphology of mammary glands in high- and low-yield camels. (<b>A</b>) Mammary glands of low-yield camels; (<b>B</b>) mammary glands of high-yield camels. AL, acinar lumen; MM, mammary mesenchyme; A, acinar epithelial cells; L, lipid droplets.</p>
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<p>Classification of identified single-nucleotide polymorphisms.</p>
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<p>Genomic regions with strong selective signals in high- and low-yielding camels. <span class="html-italic">F<sub>ST</sub></span> and <span class="html-italic">θπ</span> ratios were calculated using a sliding window analysis with a window size of 20 kb and a step size of 5 kb. Data points in blue corresponding to the top 1% of <span class="html-italic">F<sub>ST</sub></span> and the top 1% of the <span class="html-italic">θπ</span> ratio distribution are genomic regions under selection in high-yield camels. Data points in green corresponding to the top 1% of the <span class="html-italic">F<sub>ST</sub></span> and the top 1% of the <span class="html-italic">θπ</span> ratio distribution are genomic regions under selection in low-yield camels.</p>
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<p>Functional enrichment and pathway analyses. (<b>A</b>) Top 10 terms in each GO category ranked according to their statistical significance; (<b>B</b>) top 20 enriched KEGG pathways.</p>
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<p>Genotyping of Bactrian camel <span class="html-italic">ROR2</span> g.7_2434270 T &gt; A. (<b>A</b>) <span class="html-italic">ROR2</span> g.7_2434270 T &gt; A; (<b>B</b>) PCR-SSCP detection results of <span class="html-italic">ROR2</span> g.7_2434270 T &gt; A; (<b>C</b>) single peak in the sequencing peak map also indicated successful mutation creations on the <span class="html-italic">ROR2</span> g.7_2434270 T &gt; A; and (<b>D</b>) effect of <span class="html-italic">ROR2</span> g.7_2434270 genotype on milk production. * <span class="html-italic">p</span> &lt; 0.05; NS, not significant.</p>
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<p>The relationship between SNPs and milk production and milk composition traits in Bactrian camels.</p>
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<p>Geographical locations of the samples. (<b>A</b>) North-western China; (<b>B</b>) Xinjiang Altay region; (<b>C</b>) sampling camel photos.</p>
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<p>Geographical locations of the samples. (<b>A</b>) North-western China; (<b>B</b>) Xinjiang Altay region; (<b>C</b>) sampling camel photos.</p>
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14 pages, 2273 KiB  
Article
A Look Inside—Histopathological Examinations of Different Tail Tip Lesions in Dairy Cows
by Lea M. Lorenz, Marielle E. Volkwein, Christine Schmidt, Mirjam Lechner and Prisca V. Kremer-Rücker
Animals 2024, 14(14), 2094; https://doi.org/10.3390/ani14142094 - 17 Jul 2024
Viewed by 51
Abstract
Feedlot cattle are frequently affected by inflammation and necrosis of the tail tips, resulting in impeded animal welfare and economic losses. In a recent study, it was demonstrated that dairy cows are also affected by different lesions of the tail tip, including alopecia, [...] Read more.
Feedlot cattle are frequently affected by inflammation and necrosis of the tail tips, resulting in impeded animal welfare and economic losses. In a recent study, it was demonstrated that dairy cows are also affected by different lesions of the tail tip, including alopecia, annular constrictions, crusting, scaling and swelling. Despite the frequent occurrence of these lesions, the underlying etiology and pathomechanisms are unclear to date. To gain insight into this malady, we histopathologically examined 16 tail tips of slaughtered dairy cows, representing the entire range of different lesions. While macroscopically alopecic areas were characterized by the formation of granulation tissue in the dermis with an inconspicuous epidermis, we found not only dermal granulation tissue but also purulent-necrotizing inflammation with the breakdown of the basement membrane in encrusted lesions. Interestingly, in some cases, we found areas of coagulation necrosis of the epidermal and dermal layers in the crusts. Tails with macroscopical scaling were affected by ortho- and/or parakeratotic hyperkeratosis, and, in tails with macroscopical swelling, we observed a low-protein edema of the central longitudinal connective tissue of the tail. We conclude that the observed lesions might be caused by ischemia of the skin in the distal parts of the tail. Full article
(This article belongs to the Section Cattle)
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<p>Annular constrictions with alopecic but otherwise intact skin. (<b>a</b>) Distal part of a tail with two annular constrictions. The more proximal constriction encompasses about 95% of the tail circumference and the more distal constriction about 50%. The skin in these areas is alopecic but otherwise intact. (<b>b</b>) Sagittal sections through the constrictions. In both cases, the epidermis is inconspicuous. No hair follicles can be seen in the dermis underneath the constriction. (<b>c</b>) Histopathological findings of the area marked in (<b>b</b>), H.E. The epidermis (1) is without pathological findings. The dermis in the affected area is characterized by granulation tissue (2), which contains no hair follicles or sebaceous glands, in contrast to the adjacent area (3). The granulation tissue extends down to the central longitudinal connective tissue of the tail (4).</p>
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<p>Annular constrictions with crust formation. (<b>a</b>) Distal part of a tail with an annular constriction encompassing the entire circumference of the tail. The center of the constriction is bloody encrusted. (<b>b</b>) Sagittal section through the annular constriction shown in (<b>a</b>). In the marginal area, the skin is alopecic and no hair follicles or sebaceous glands can be found in the area under the constriction. In the central area of the constriction, there are bloody crusts. (<b>c</b>) Distal part of a tail with annular constriction encompassing about 70% of the tail. The constriction is bloody encrusted. The tail tip is also affected by severe scaling. (<b>d</b>) Distal part of a tail with two annular constrictions with crust formation. The more proximal constriction encompasses about 75% of the tail circumference. (<b>e</b>) Sagittal section of the proximal constriction shown in (<b>d</b>). The constricted area is devoid of hair follicles or sebaceous glands. In the center of the constriction, the epidermis is covered by crusts.</p>
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<p>Histological sections of annular constrictions with crust formation, H.E. (<b>a</b>) Histological section of the constricted area marked in 2 (<b>a</b>) and 2 (<b>b</b>). Purulent-necrotizing inflammation of the epidermis with crusting (1), breakdown of the epidermal basement membrane (2) and underlying granulation tissue (3). (<b>b</b>) Magnification of the granulation tissue depicted in (<b>a</b>) directly underlying the lesion. Accumulation of mononuclear cells (1) and sprouting new blood vessels (2). (<b>c</b>) Magnification of the granulation tissue depicted in (<b>a</b>) in the deeper layers underneath the lesion, containing blood vessels (1), fibrocytes (2) and collagen (3). (<b>d</b>) Magnification of the area marked in (<b>a</b>) with spongiosis (1), hemorrhage (2) and the breakdown of the basement membrane (3). (<b>e</b>) Histological section of the constricted area marked in 2 (<b>c</b>), showing the accumulation of neutrophil granulocytes (1), parakeratotic hyperkeratosis (2) and intracellular edema of keratinocytes (3). Dermis displays hyperemia of papillary bodies (4) and adjacent free erythrocytes (arrows). (<b>f</b>) Histological section of the constricted area marked in 2 (<b>d</b>) and 2 (<b>e</b>). The crust contains neutrophil granulocytes (1), bleeding (2) and intralesional bacteria (arrow). (<b>g</b>) Magnification of the area marked in (<b>f</b>). Coagulation necrosis of the dermal papillary bodies (1) and epidermal layers within the crust. Necrotic area is demarcated by neutrophil granulocytes (2). (<b>h</b>) Magnification of the necrotic area depicted in (<b>g</b>). Structure of the papillary bodies is still intact with visible vessel walls (arrows). Cells of the dermal (1) and epidermal layers (2) are necrotic.</p>
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<p>Alopecic tail with encrustation of the tail tip. (<b>a</b>) Distal part of a tail with large-scale alopecia and annular constriction encompassing about 95% of the tail circumference. (<b>b</b>) Tip of the tail depicted in (<b>a</b>) with alopecia and encrustation. (<b>c</b>) Sagittal section of the same tail with loss of tissue at the tip of the tail. (<b>d</b>) Histological section of the area marked in (<b>c</b>) with purulent-necrotizing inflammation of the epidermis and crust formation showing the accumulation of neutrophil granulocytes (1) and the multifocal breakdown of the basement membrane (2).</p>
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<p>Tails with scaling, swelling or wart-like masses. (<b>a</b>) Distal part of a tail with severe scaling. (<b>b</b>) Histological section of the tail tip depicted in (<b>a</b>), H.E. Epidermis displays orthokeratotic hyperkeratosis (1). (<b>c</b>) Distal part of a tail with macroscopically visible and palpable swelling. (<b>d</b>) Sagittal section of the tail shown in (<b>c</b>). (<b>e</b>) Histological section of the area marked in (<b>d</b>), H.E. Extracellular, low-protein edema (arrows) in the region of the central longitudinal arranged connective tissue. (<b>f</b>) Distal part of a tail with single wart-like mass. (<b>g</b>) Histological section of the wart-like mass shown in (<b>f</b>), H.E. Inconspicuous epidermis (1) with underlying layer of fibroblasts (arrow), connective tissue matrix (2) and blood vessels (3).</p>
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23 pages, 882 KiB  
Review
The Assessment of Sperm DNA Integrity: Implications for Assisted Reproductive Technology Fertility Outcomes across Livestock Species
by Maya J. Robertson, Caitlin Chambers, Eloise A. Spanner, Simon P. de Graaf and Jessica P. Rickard
Biology 2024, 13(7), 539; https://doi.org/10.3390/biology13070539 (registering DOI) - 17 Jul 2024
Viewed by 63
Abstract
Sperm DNA integrity is increasingly considered a useful measure of semen quality in mammalian reproduction. However, the definition of DNA integrity, the ideal means by which it should be measured, and its predictive value for fertility remain a topic of much discussion. With [...] Read more.
Sperm DNA integrity is increasingly considered a useful measure of semen quality in mammalian reproduction. However, the definition of DNA integrity, the ideal means by which it should be measured, and its predictive value for fertility remain a topic of much discussion. With an emphasis on livestock species, this review discusses the assays that have been developed to measure DNA integrity as well as their correlation with in vitro and in vivo fertility. Full article
(This article belongs to the Section Zoology)
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<p>An overview of sperm development and chromatin dynamics during histone–protamine exchange. (<b>A</b>) Spermatocytes undergo mitotic division to form haploid cells, which then undergo meiosis to form spermatids. Ejaculated sperm can be collected for in vitro processing or be deposited into the female reproductive tract, where they undergo the capacitation and acrosome reaction [<a href="#B21-biology-13-00539" class="html-bibr">21</a>], enabling penetration of the oocyte zone pellucida via the acrosome. (<b>B</b>) As chromatin compaction progresses during testicular development and epididymal maturation, the nucleosomal architecture shifts to a toroidal structure facilitated by sperm nuclear basic proteins. During spermatid elongation, transitional proteins are expressed, leading to histone replacement by protamine proteins. This results in tightly packaged toroids of DNA attached to the sperm nuclear MAR stacked side by side with a small number of retained histones. Following fertilisation, sperm-specific protamines are replaced with oocyte-supplied histones. Histone-bound chromatin and MARs from the sperm are retained in the newly formed pronucleus, altering chromatin accessibility. The yellow boxes denote potential causes of DNA damage or embryo development disruptions throughout these processes. Figure created with BioRender.com. This protamine-bound chromatin structure is a model of the most efficient form toroids could condense to, based on a figure published by Ward, 2009 [<a href="#B23-biology-13-00539" class="html-bibr">23</a>]. Abbreviations: DNA, deoxynucleic acid; MAR, matrix attachment region.</p>
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20 pages, 4712 KiB  
Article
X-ray Microtomography Analysis of Integrated Crop–Livestock Production’s Impact on Soil Pore Architecture
by José V. Gaspareto and Luiz F. Pires
AgriEngineering 2024, 6(3), 2249-2268; https://doi.org/10.3390/agriengineering6030132 (registering DOI) - 17 Jul 2024
Viewed by 81
Abstract
Integrated crop–livestock production (ILP) is an interesting alternative for more sustainable soil use. However, more studies are needed to analyze the soil pore properties under ILP at the micrometer scale. Thus, this study proposes a detailed analysis of the soil pore architecture at [...] Read more.
Integrated crop–livestock production (ILP) is an interesting alternative for more sustainable soil use. However, more studies are needed to analyze the soil pore properties under ILP at the micrometer scale. Thus, this study proposes a detailed analysis of the soil pore architecture at the micrometer scale in three dimensions. For this purpose, samples of an Oxisol under ILP subjected to minimum tillage (MT) and no tillage (NT) with ryegrass as the cover crop (C) and silage (S) were studied. The micromorphological properties of the soil were analyzed via X-ray microtomography. The MT(C) system showed the highest values of porosity (c. 20.4%), connectivity (c. 32.8 × 103), volume (c. 26%), and the number of pores (c. 32%) in a rod-like shape. However, the MT(S), NT(C), and NT(S) systems showed greater tortuosity (c. 2.2, c. 2.0, and c. 2.1) and lower pore connectivity (c. 8.3 × 103, c. 6.9 × 103, and c. 6.2 × 103), especially in S use. Ellipsoidal and rod-shaped pores predominated over spheroidal and disc-shaped pores in all treatments. The results of this study show that the use of ryegrass as a cover crop improves the soil physical properties, especially in MT. For S use, the type of soil management (MT or NT) did not show any differences. Full article
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<p>Map of the study region showing the state of Paraná, the municipality of Castro and the experimental area. The region highlighted in red represents the experimental plot studied.</p>
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<p>Procedures for image analysis using X-ray tomography: (<b>a</b>) selection of the region of interest (ROI) inside the scanned aggregate (area indicated in yellow); (<b>b</b>) ROI from the clipping procedure; (<b>c</b>) application of the median 3D filter; (<b>d</b>) application of the unsharp mask tool; (<b>e</b>) segmentation by Otsu method; (<b>f</b>) noise removal.</p>
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<p>Flowchart showing the main steps carried out in our study to measure the morphological and geometric properties of the soil.</p>
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<p>Three-dimensional (3D) images—reconstructed by X-ray microtomography of the different soil management systems (minimum tillage + ryegrass cover, MT(C); minimum tillage + ryegrass silage, MT(S); no tillage + ryegrass cover, NT(C); and no tillage + ryegrass silage, NT(S)). In the images on the left, the pores are shown in black and the soil matrix in gray. In the images on the right, the pores are shown in terracotta.</p>
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<p>Morphological and geometric soil properties for the minimum tillage system (MT), no tillage system (NT), and ryegrass (C: cover crop; S: silage). (<b>a</b>) Imaged porosity (<math display="inline"><semantics> <mrow> <mi>φ</mi> </mrow> </semantics></math>), (<b>b</b>) fractal dimension (<math display="inline"><semantics> <mrow> <mi>F</mi> <mi>D</mi> </mrow> </semantics></math>), (<b>c</b>) degree of anisotropy (<math display="inline"><semantics> <mrow> <mi>D</mi> </mrow> </semantics></math>A), and (<b>d</b>) pore connectivity (<math display="inline"><semantics> <mrow> <mi>C</mi> </mrow> </semantics></math>). Different letters indicate significant differences between treatments (<span class="html-italic">p</span> &lt; 0.05). The error bars (red lines) indicate the standard deviation of the mean.</p>
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<p>Tortuosity (<math display="inline"><semantics> <mrow> <mi mathvariant="normal">Ʈ</mi> </mrow> </semantics></math>) calculated based on 3D images for the minimum tillage system (MT), no tillage system (NT), and ryegrass (C: cover; S: silage). (<b>a</b>) Tortuosity in the <math display="inline"><semantics> <mrow> <mi>x</mi> </mrow> </semantics></math> direction (<math display="inline"><semantics> <mrow> <mi mathvariant="normal">Ʈ</mi> <mi>x</mi> </mrow> </semantics></math>), (<b>b</b>) tortuosity in the <math display="inline"><semantics> <mrow> <mi>y</mi> </mrow> </semantics></math> direction (<math display="inline"><semantics> <mrow> <mi mathvariant="normal">Ʈ</mi> <mi>y</mi> </mrow> </semantics></math>), (<b>c</b>) tortuosity in the <math display="inline"><semantics> <mrow> <mi>z</mi> </mrow> </semantics></math> direction (<math display="inline"><semantics> <mrow> <mi mathvariant="normal">Ʈ</mi> <mi>z</mi> </mrow> </semantics></math>), and (<b>d</b>) tortuosity (<math display="inline"><semantics> <mrow> <mi mathvariant="normal">Ʈ</mi> </mrow> </semantics></math>) considering all directions. Different letters indicate significant differences between treatments (<span class="html-italic">p</span> &lt; 0.05). The error bars (red lines) indicate the standard deviation of the mean.</p>
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<p>Contribution of volume (<span class="html-italic">VP</span>) and number of pores (<span class="html-italic">NP</span>) in relation to total volume for the different pore shapes (equant—spheroidal shape, prolate—rod-like shape, oblate—disk-like shape, and triaxial—ellipsoidal shape) for the following treatments: minimum tillage system (MT), no tillage system (NT), and ryegrass (C: cover; S: silage). (<b>a</b>) Volume of pores (<span class="html-italic">VP</span>) and (<b>b</b>) number of pores (<span class="html-italic">NP</span>). Different letters indicate significant differences between treatments (<span class="html-italic">p</span> &lt; 0.05). The error bars (red lines) indicate the standard deviation of the mean.</p>
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<p>Scores of PC1 and PC2 for soil samples under different treatments (minimum tillage + ryegrass cover, MT(C); minimum tillage + ryegrass silage, MT(S); no tillage + ryegrass cover, NT(C); and no tillage + ryegrass silage, NT(S)) with the inclusion of the following variables: pore connectivity (<math display="inline"><semantics> <mrow> <mi>C</mi> </mrow> </semantics></math>), porosity (<math display="inline"><semantics> <mrow> <mi>φ</mi> </mrow> </semantics></math>), tortuosity (<math display="inline"><semantics> <mrow> <mi mathvariant="normal">Ʈ</mi> </mrow> </semantics></math>), fractal dimension (<math display="inline"><semantics> <mrow> <mi>F</mi> <mi>D</mi> </mrow> </semantics></math>), degree of anisotropy (<math display="inline"><semantics> <mrow> <mi>D</mi> <mi>A</mi> </mrow> </semantics></math>), volume of pores (<math display="inline"><semantics> <mrow> <mi>V</mi> <mi>P</mi> </mrow> </semantics></math>), number of pores (<math display="inline"><semantics> <mrow> <mi>N</mi> <mi>P</mi> </mrow> </semantics></math>), and shape of pores (equant—spheroidal shape, prolate—rod-like shape, oblate—disk-like shape, and triaxial—ellipsoidal shape).</p>
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25 pages, 2892 KiB  
Article
Ratanjot (Alkanna tinctoria L.) Root Extract, Rich in Antioxidants, Exhibits Strong Antimicrobial Activity against Foodborne Pathogens and Is a Potential Food Preservative
by Annada Das, Subhasish Biswas, Kaushik Satyaprakash, Dipanwita Bhattacharya, Pramod Kumar Nanda, Gopal Patra, Sushmita Moirangthem, Santanu Nath, Pubali Dhar, Arun K. Verma, Olipriya Biswas, Nicole Irizarry Tardi, Arun K. Bhunia and Arun K. Das
Foods 2024, 13(14), 2254; https://doi.org/10.3390/foods13142254 - 17 Jul 2024
Viewed by 99
Abstract
Natural and sustainable plant-based antioxidants and antimicrobials are highly desirable for improving food quality and safety. The present investigation assessed the antimicrobial and antioxidant properties of active components from Alkanna tinctoria L. (herb) roots, also known as Ratanjot root. Two methods were used [...] Read more.
Natural and sustainable plant-based antioxidants and antimicrobials are highly desirable for improving food quality and safety. The present investigation assessed the antimicrobial and antioxidant properties of active components from Alkanna tinctoria L. (herb) roots, also known as Ratanjot root. Two methods were used to extract active components: microwave-assisted hot water (MAHW) and ethanolic extraction. MAHW extract yielded 6.29%, while the ethanol extract yielded 18.27%, suggesting superior Ratanjot root extract powder (RRP) solubility in ethanol over water. The ethanol extract showed significantly higher antioxidant activity than the MAHW extract. Gas Chromatography–Mass Spectrometry analysis revealed three major phenolic compounds: butanoic acid, 3-hydroxy-3-methyl-; arnebin 7, and diisooctyl pthalate. The color attributes (L*, a*, b*, H°ab, C*ab) for the ethanolic and MAHW extracts revealed significant differences (p < 0.05) in all the above parameters for both types of extracts, except for yellowness (b*) and chroma (C*ab) values. The ethanol extract exhibited antimicrobial activity against 14 foodborne bacteria, with a significantly higher inhibitory effect against Gram-positive bacteria (Listeria monocytogenes and Staphylococcus aureus) than the Gram-negative bacteria (Salmonella enterica serovar Typhimurium and Escherichia coli). The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were both 25 mg/mL for the Gram-negative bacteria, while the MIC and MBC concentrations varied for Gram-positive bacteria (0.049–0.098 mg/mL and 0.098–0.195 mg/mL) and the antimicrobial effect was bactericidal. The antimicrobial activities of RRP extract remained stable under broad temperature (37–100 °C) and pH (2–6) conditions, as well as during refrigerated storage for 30 days. Application of RRP at 1% (10 mg/g) and 2.5% (25 mg/g) levels in a cooked chicken meatball model system prevented lipid oxidation and improved sensory attributes and retarded microbial growth during refrigerated (4 °C) storage for 20 days. Furthermore, the RRP extract was non-toxic when tested with sheep erythrocytes and did not inhibit the growth of probiotics, Lacticaseibacillus casei, and Lactiplantibacillus plantarum. In conclusion, the study suggests that RRP possesses excellent antimicrobial and antioxidant activities, thus making it suitable for food preservation. Full article
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<p><span class="html-italic">Alkanna tinctoria</span> (known as Ratanjot) plant and roots.</p>
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<p>Antioxidant activity of ethanol (EE) and water extract (MAHW) of Ratanjot (<span class="html-italic">Alkanna tinctoria</span> L.) root (RRP) extract. Bars are average (n = 6) ± SE bearing different superscript(s) (A, B, C, D) differ significantly at <span class="html-italic">p</span> &lt; 0.05. TPC, total phenolic compound; ABTS, 2.2-Azino-bis (3-ethylbenzothiazoline-6-sulfonic acid); DPPH, 2,2-Diphenyl-1-picrylhydrazyl (DPPH); GAE, gallic acid equivalent.</p>
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<p>Gas chromatography and mass spectrometry analysis of <span class="html-italic">Alkanna tinctoria</span> L. (known as Ratanjot) root ethanol extract produced three major components: Butanoic acid, 3-hydroxy-3-methyl-; Arnebin 7, and Diisooctyl pthalate.</p>
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<p>Growth kinetics of (<b>A</b>) <span class="html-italic">Staphylococcus aureus</span> (SA) ATCC 25923 and (<b>B</b>) <span class="html-italic">Listeria monocytogenes</span> (LM) ATCC 19111 after exposure to Ratanjot (<span class="html-italic">Alkanna tinctoria</span> L.) root extract (ethanol extract), RRP at 1X MIC and MBC for 24 h. Ciprofloxacin (1 X MIC and 1 X MBC) was used as a positive control.</p>
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<p>Antimicrobial activity of Ratanjot (<span class="html-italic">Alkanna tinctoria</span> L.) root extract (RRP) exposed to various (<b>A</b>) pH ranges 2–8, (<b>B</b>) temperature ranges (37–100 °C) and (<b>C</b>) Storage days (0–30 days) at 4 °C for 30 days against <span class="html-italic">Listeria monocytogenes</span> and <span class="html-italic">Staphylococcus aureus</span> analyzed by agar well-diffusion assay. Bars marked with different letters (a, b, c) are significantly different at <span class="html-italic">p</span> &lt; 0.05. UD: Undetected.</p>
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<p>Effect of Ratanjot (<span class="html-italic">Alkanna tinctoria</span> L.) root extract (RRP) at 1X and 2X MIC on <span class="html-italic">Lactobacillus</span> growth. (<b>A</b>) <span class="html-italic">Lactiplantibacillus plantarum</span> and <span class="html-italic">Lacticaseibacillus casei</span> counts (log 10 CFU/mL) after 24 h in MRS broth, (<b>B</b>) visual examination of turbidity of <span class="html-italic">L. casei</span> growth after 24 h.</p>
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<p>In vitro hemolysis assay of Ratanjot (<span class="html-italic">Alkanna tinctoria</span> L.) root extract (RRP) at various concentrations (1–10 XMIC). (<b>A</b>) Bar diagram showing percent hemolysis of sheep red blood cells. Data are mean (n = 6) ± SE. *, <span class="html-italic">p</span> &lt; 0.05. Triton X-100 (0.1%) used as a positive control for complete lysis. (<b>B</b>) Photographs showing the difference in hemolysis and no hemolysis of RBC.</p>
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<p>Application of Ratanjot (<span class="html-italic">Alkanna tinctoria</span> L.) root extract (RRP) at 1% (T1) and 2.5% (T2) in chicken meatballs on antioxidant (TBARS) and antimicrobial activity stored at 4 °C for 20 days. (<b>A</b>) Chicken meat balls incorporated with RRP. (<b>B</b>) TBARS values, (<b>C</b>) TVC, (<b>D</b>) Psychrophilic counts, (<b>E</b>) <span class="html-italic">E. coli</span> counts, and (<b>F</b>) <span class="html-italic">S. aureus</span> counts of control and RRP incorporated meat balls during refrigerated storage for 20 days. Bars marked with uppercase letters (A, B, C, D, E) signify differences in values on various days, and lowercase letters (a, b, c) signify differences within the same day for all three samples. <span class="html-italic">p</span> &lt; 0.05. UD: Undetected.</p>
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15 pages, 2523 KiB  
Article
Chromosome-Scale Genome Assembly of the Sheep-Biting Louse Bovicola ovis Using Nanopore Sequencing Data and Pore-C Analysis
by Chian Teng Ong, Karishma T. Mody, Antonino S. Cavallaro, Yakun Yan, Loan T. Nguyen, Renfu Shao, Neena Mitter, Timothy J. Mahony and Elizabeth M. Ross
Int. J. Mol. Sci. 2024, 25(14), 7824; https://doi.org/10.3390/ijms25147824 - 17 Jul 2024
Viewed by 121
Abstract
Bovicola ovis, commonly known as the sheep-biting louse, is an ectoparasite that adversely affects the sheep industry. Sheep louse infestation lowers the quality of products, including wool and leather, causing a loss of approximately AUD 123M per annum in Australia alone. The [...] Read more.
Bovicola ovis, commonly known as the sheep-biting louse, is an ectoparasite that adversely affects the sheep industry. Sheep louse infestation lowers the quality of products, including wool and leather, causing a loss of approximately AUD 123M per annum in Australia alone. The lack of a high-quality genome assembly for the sheep-biting louse, as well as any closely related livestock lice, has hindered the development of louse research and management control tools. In this study, we present the assembly of B. ovis with a genome size of ~123 Mbp based on a nanopore long-read sequencing library and Illumina RNA sequencing, complemented with a chromosome-level scaffolding using the Pore-C multiway chromatin contact dataset. Combining multiple alignment and gene prediction tools, a comprehensive annotation on the assembled B. ovis genome was conducted and recalled 11,810 genes as well as other genomic features including orf, ssr, rRNA and tRNA. A manual curation using alignment with the available closely related louse species, Pediculus humanus, increased the number of annotated genes to 16,024. Overall, this study reported critical genetic resources and biological insights for the advancement of sheep louse research and the development of sustainable control strategies in the sheep industry. Full article
(This article belongs to the Special Issue Regulation by Non-coding RNAs 2024)
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<p>Schematic representation of the <span class="html-italic">B. ovis</span> genome statistics: (<b>A</b>) Distribution of k-mer frequency in <span class="html-italic">B. ovis</span> sequencing data generated with Nanopore sequencing technology at k-mer size of 21. This GenomeScope profile describes the estimated length (len), percent of the genome that is unique (uniq), rate of homozygosity (aa), rate of heterozygosity (ab), mean k-mer coverage (kcov), error rate (err) and average rate of duplication (dup). (<b>B</b>) The <span class="html-italic">B. ovis</span> genome contig contact matrix using pore-C data. The colour intensity represents the Pore-C contact density in the plot.</p>
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<p>The sheep-biting louse <span class="html-italic">B. ovis</span> genome. The circus plot shows the genomic features for 13 pseudo-chromosomes: (A) Pseudo-chromosome, (B) GC content, (C) GC skew and (D) gene density. Gene density was represented by a colour gradient ranging from red to yellow, with higher density indicated by red, and lower density indicated by yellow. The pseudo-chromosome size scale is shown in Mb scale.</p>
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<p>The COG functional annotation of the uniquely expressed genes in <span class="html-italic">B. ovis</span>. COG groups with less than 4% were grouped as “Others”, the details of these COG groups are listed in <a href="#app1-ijms-25-07824" class="html-app">Supplementary File S1</a>.</p>
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20 pages, 4689 KiB  
Article
Extending Multi-Output Methods for Long-Term Aboveground Biomass Time Series Forecasting Using Convolutional Neural Networks
by Efrain Noa-Yarasca, Javier M. Osorio Leyton and Jay P. Angerer
Mach. Learn. Knowl. Extr. 2024, 6(3), 1633-1652; https://doi.org/10.3390/make6030079 (registering DOI) - 17 Jul 2024
Viewed by 119
Abstract
Accurate aboveground vegetation biomass forecasting is essential for livestock management, climate impact assessments, and ecosystem health. While artificial intelligence (AI) techniques have advanced time series forecasting, a research gap in predicting aboveground biomass time series beyond single values persists. This study introduces RECMO [...] Read more.
Accurate aboveground vegetation biomass forecasting is essential for livestock management, climate impact assessments, and ecosystem health. While artificial intelligence (AI) techniques have advanced time series forecasting, a research gap in predicting aboveground biomass time series beyond single values persists. This study introduces RECMO and DirRecMO, two multi-output methods for forecasting aboveground vegetation biomass. Using convolutional neural networks, their efficacy is evaluated across short-, medium-, and long-term horizons on six Kenyan grassland biomass datasets, and compared with that of existing single-output methods (Recursive, Direct, and DirRec) and multi-output methods (MIMO and DIRMO). The results indicate that single-output methods are superior for short-term predictions, while both single-output and multi-output methods exhibit a comparable effectiveness in long-term forecasts. RECMO and DirRecMO outperform established multi-output methods, demonstrating a promising potential for biomass forecasting. This study underscores the significant impact of multi-output size on forecast accuracy, highlighting the need for optimal size adjustments and showcasing the proposed methods’ flexibility in long-term forecasts. Short-term predictions show less significant differences among methods, complicating the identification of the best performer. However, clear distinctions emerge in medium- and long-term forecasts, underscoring the greater importance of method choice for long-term predictions. Moreover, as the forecast horizon extends, errors escalate across all methods, reflecting the challenges of predicting distant future periods. This study suggests advancing hybrid models (e.g., RECMO and DirRecMO) to improve extended horizon forecasting. Future research should enhance adaptability, investigate multi-output impacts, and conduct comparative studies across diverse domains, datasets, and AI algorithms for robust insights. Full article
(This article belongs to the Section Network)
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<p>Time series of aboveground vegetation biomass (kg/ha) derived from calibrated PHYGROW model simulations at six rangeland locations in Kenya. Each subfigure (<b>a</b>–<b>f</b>) represents time series data from a representative location.</p>
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<p>Conversion of time series data into a supervised dataset. Blue squares: predictor subsequences inputted to the model (window size w). Orange squares: predicted values.</p>
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<p>Overview of the examined and prospective forecasting methods. Blue squares: predictor subsequences inputted to the model (window size w). Orange squares: predicted values.</p>
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<p>Architecture of the Convolutional Neural Network.</p>
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<p>Average RMSE values across different forecasting methods for aboveground vegetation biomass (kg/ha) across a horizon (<span class="html-italic">H</span>) of 24. Each subfigure from (<b>a</b>–<b>f</b>) displays the results of each time series data from a representative study location.</p>
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<p>Relative RMSE values of aboveground vegetation biomass (kg/ha) across different forecasting methods in relation to forecast horizons. Relative RMSE is calculated as a percentage by dividing the average RMSE by the time series mean.</p>
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16 pages, 7075 KiB  
Article
Genomic Epidemiology of Rift Valley Fever Virus Involved in the 2018 and 2022 Outbreaks in Livestock in Rwanda
by Isidore Nsengimana, John Juma, Kristina Roesel, Methode N. Gasana, Fabrice Ndayisenga, Claude M. Muvunyi, Emmanuel Hakizimana, Jean N. Hakizimana, Gillian Eastwood, Augustino A. Chengula, Bernard Bett, Christopher J. Kasanga and Samuel O. Oyola
Viruses 2024, 16(7), 1148; https://doi.org/10.3390/v16071148 - 17 Jul 2024
Viewed by 205
Abstract
Rift Valley fever (RVF), a mosquito-borne transboundary zoonosis, was first confirmed in Rwanda’s livestock in 2012 and since then sporadic cases have been reported almost every year. In 2018, the country experienced its first large outbreak, which was followed by a second one [...] Read more.
Rift Valley fever (RVF), a mosquito-borne transboundary zoonosis, was first confirmed in Rwanda’s livestock in 2012 and since then sporadic cases have been reported almost every year. In 2018, the country experienced its first large outbreak, which was followed by a second one in 2022. To determine the circulating virus lineages and their ancestral origin, two genome sequences from the 2018 outbreak, and thirty-six, forty-one, and thirty-eight sequences of small (S), medium (M), and large (L) genome segments, respectively, from the 2022 outbreak were generated. All of the samples from the 2022 outbreak were collected from slaughterhouses. Both maximum likelihood and Bayesian-based phylogenetic analyses were performed. The findings showed that RVF viruses belonging to a single lineage, C, were circulating during the two outbreaks, and shared a recent common ancestor with RVF viruses isolated in Uganda between 2016 and 2019, and were also linked to the 2006/2007 largest East Africa RVF outbreak reported in Kenya, Tanzania, and Somalia. Alongside the wild-type viruses, genetic evidence of the RVFV Clone 13 vaccine strain was found in slaughterhouse animals, demonstrating a possible occupational risk of exposure with unknown outcome for people working in meat-related industry. These results provide additional evidence of the ongoing wide spread of RVFV lineage C in Africa and emphasize the need for an effective national and international One Health-based collaborative approach in responding to RVF emergencies. Full article
(This article belongs to the Special Issue Emerging Highlights in the Study of Rift Valley Fever Virus)
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<p>Administrative map of Rwanda, showing in red stars the location of RVF emergency testing centers where slaughterhouse samples used in this study were screened during the 2022 Rwanda RVF outbreak. The green, blue, and pink colors show the Kigali, Huye and Nyamasheke RVF testing center catchment areas, respectively. The dark brown color indicates collection sites for RVFV successfully sequenced for the 2018 outbreak. The map was drawn using QGIS version 3.24.1, freely available at <a href="https://www.qgis.org/en/site/" target="_blank">https://www.qgis.org/en/site/</a>, accessed on 25 May 2024.</p>
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<p>Bayesian molecular clock analysis of RVFV strains based on L segment. The time-scaled maximum clade credibility tree is depicted with branches colored according to country of origin of the viruses analyzed. The phylogenetic tree tips are labeled as accession, name, country, and year of collection of the specimens. RVFV Clone 13 sequences isolated in Rwanda are shown in the box. The letters indicate RVFV lineages. Posterior support values (HPD) are shown above each node (i.e., 1 = 100% support).</p>
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<p>Bayesian molecular clock analysis of RVFV strains based on M segment. The time-scaled maximum clade credibility tree is shown with branches colored according to country of origin of the viruses analyzed. The phylogenetic tree tips are labeled as accession, name, country, and year of collection of the specimens. RVFV Clone 13 sequences isolated in Rwanda are shown in the box. The letters indicate RVFV lineages. Posterior support values (HPD) are shown above each node (i.e., 1 = 100% support).</p>
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<p>Bayesian molecular clock analysis of RVFV strains based on S segment. The time-scaled maximum clade credibility tree is presented with branches colored according to country of origin of the viruses analyzed. The phylogenetic tree tips are labeled as accession, name, country, and year of collection of the specimens. RVFV Clone 13 sequences isolated in Rwanda are shown in the box. The letters indicate RVFV lineages. Posterior support values (HPD) are shown above each node (i.e., 1 = 100% support).</p>
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9 pages, 575 KiB  
Article
New Insights into the Phylogeny of the A.Br.161 (“A.Br.Heroin”) Clade of Bacillus anthracis
by Markus Antwerpen, Wolfgang Beyer and Gregor Grass
Pathogens 2024, 13(7), 593; https://doi.org/10.3390/pathogens13070593 - 16 Jul 2024
Viewed by 191
Abstract
Bacillus anthracis is a rare but highly dangerous zoonotic bacterial pathogen. At the beginning of this century, a new manifestation of the disease, injectional anthrax, emerged as a result of recreational heroin consumption involving contaminated drugs. The organisms associated with this 13-year-lasting outbreak [...] Read more.
Bacillus anthracis is a rare but highly dangerous zoonotic bacterial pathogen. At the beginning of this century, a new manifestation of the disease, injectional anthrax, emerged as a result of recreational heroin consumption involving contaminated drugs. The organisms associated with this 13-year-lasting outbreak event in European drug consumers were all grouped into the canonical single-nucleotide polymorphism (canSNP) clade A-branch (A.Br.) 161 of B. anthracis. Related clade A.Br.161 strains of B. anthracis not associated with heroin consumption have also been identified from different countries, mostly in Asia. Because of inadvertent spread by anthropogenic activities, other strains of this A.Br.161 lineage were, however, isolated from several countries. Thus, without additional isolates from this clade, its origin of evolution or its autochthonous region remains obscure. Here, we genomically characterized six new A.Br.161 group isolates, some of which were from Iran, with others likely historically introduced into Germany. All the chromosomes of these isolates could be grouped into a distinct sub-clade within the A.Br.161 clade. This sub-clade is separated from the main A.Br.161 lineage by a single SNP. We have developed this SNP into a PCR assay facilitating the future attribution of strains to this group. Full article
(This article belongs to the Special Issue Anthrax—a Threat beyond Bacillus anthracis)
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<p>Phylogenetic position and minimum spanning tree (MST) of canonical A.Br.161 clade of <span class="html-italic">B. anthracis</span>. (<b>left</b>) Overview of phylogenetic placement of canSNP clade A. Br.161 within the global diversity of <span class="html-italic">B. anthracis</span>; (<b>right</b>) A MST was calculated from 1550 chromosomal SNPs and visualized using Grapetree [<a href="#B24-pathogens-13-00593" class="html-bibr">24</a>]. Numerical SNP distances between chromosomes (respective strains are named in bold-type letters) are indicated at branches. Newly sequenced isolate names are displayed in red, sub-clades in blue, and relevant SNP positions in green characters. Countries from which respective strains were isolated are ISO 3166-1 alpha-2 coded as grey letters: CN—China, DE—Germany, DK—Denmark, ET—Ethiopia, FR—France, IR—Iran, KZ—Kazakhstan, NO—Norway, PK—Pakistan, TR—Turkey, UK—United Kingdom, US—United States of America.</p>
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19 pages, 7197 KiB  
Article
Evaluation of the Efficiency of Machine Learning Algorithms for Identification of Cattle Behavior Using Accelerometer and Gyroscope Data
by Tsvetelina Mladenova, Irena Valova, Boris Evstatiev, Nikolay Valov, Ivan Varlyakov, Tsvetan Markov, Svetoslava Stoycheva, Lora Mondeshka and Nikolay Markov
AgriEngineering 2024, 6(3), 2179-2197; https://doi.org/10.3390/agriengineering6030128 (registering DOI) - 16 Jul 2024
Viewed by 204
Abstract
Animal welfare is a daily concern for livestock farmers. It is known that the activity of cows characterizes their general physiological state and deviations from the normal parameters could be an indicator of different kinds of diseases and conditions. This pilot study investigated [...] Read more.
Animal welfare is a daily concern for livestock farmers. It is known that the activity of cows characterizes their general physiological state and deviations from the normal parameters could be an indicator of different kinds of diseases and conditions. This pilot study investigated the application of machine learning for identifying the behavioral activity of cows using a collar-mounted gyroscope sensor and compared the results with the classical accelerometer approach. The sensor data were classified into three categories, describing the behavior of the animals: “standing and eating”, “standing and ruminating”, and “laying and ruminating”. Four classification algorithms were considered—random forest ensemble (RFE), decision trees (DT), support vector machines (SVM), and naïve Bayes (NB). The training relied on manually classified data with a total duration of 6 h, which were grouped into 1s, 3s, and 5s piles. The obtained results showed that the RFE and DT algorithms performed the best. When using the accelerometer data, the obtained overall accuracy reached 88%; and when using the gyroscope data, the obtained overall accuracy reached 99%. To the best of our knowledge, no other authors have previously reported such results with a gyroscope sensor, which is the main novelty of this study. Full article
(This article belongs to the Section Livestock Farming Technology)
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<p>General schematic of the experimental setup.</p>
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<p>A cow with a collar and an experimental IoT device (<b>a</b>), and closeup of the experimental IoT module (<b>b</b>).</p>
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<p>Conceptual diagram of the data collection and analysis system.</p>
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<p>A sample fragment from the training database.</p>
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<p>Adapted classification methodology in this study.</p>
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<p>Distribution of the accelerometer data before (<b>a</b>–<b>c</b>) and after (<b>d</b>–<b>f</b>) the outlier detection algorithm was applied.</p>
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<p>Distribution of the gyroscope data before (<b>a</b>–<b>c</b>) and after (<b>d</b>–<b>f</b>) the outlier detection algorithm was applied.</p>
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<p>Class distribution for the accelerometer and gyroscope datasets with 1s piles.</p>
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<p>Distribution of the gyroscope data grouped in 5s piles.</p>
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<p>Comparison of the overall accuracy results for the different training algorithms, datasets, and grouping intervals.</p>
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<p>F1 Score for each class individually.</p>
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16 pages, 4742 KiB  
Article
Isolation, Characterization, and Functional Properties of Antioxidant Peptides from Mulberry Leaf Enzymatic Hydrolysates
by Yichen Zhou, Rijun Zhang, Junyong Wang, Yucui Tong, Jing Zhang, Zhenzhen Li, Haosen Zhang, Zaheer Abbas, Dayong Si and Xubiao Wei
Antioxidants 2024, 13(7), 854; https://doi.org/10.3390/antiox13070854 - 16 Jul 2024
Viewed by 191
Abstract
Recent evidence suggests that mulberry leaves have good antioxidant activity. However, what the antioxidant ingredient is and how the ingredient works are still not well understood. In this study, we enzymatically hydrolyze mulberry leaf proteins (MLPs) using neutral protease and find that the [...] Read more.
Recent evidence suggests that mulberry leaves have good antioxidant activity. However, what the antioxidant ingredient is and how the ingredient works are still not well understood. In this study, we enzymatically hydrolyze mulberry leaf proteins (MLPs) using neutral protease and find that the mulberry leaf protein hydrolysates (MLPHs) have stronger antioxidant activity compared to MLPs. We separate the core antioxidant components in MLPHs by ion-exchange columns and molecular sieves and identify 798 antioxidant peptides by LC-MS/MS. Through bioinformatics analysis and biochemical assays, we screen two previously unreported peptides, P6 and P7, with excellent antioxidant activities. P6 and P7 not only significantly reduce ROS in cells but also improve the activities of the antioxidant enzymes SOD and CAT. In addition, both peptides are found to exert protective effects against H2O2-induced chromatin damage and cell apoptosis. Collectively, these results provide support for the application of mulberry leaf peptides as antioxidants in the medical, food and livestock industries. Full article
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<p>Antioxidant capacities of mulberry leaf protein (MLP) and mulberry leaf protein hydrolysates (MLPHs). (<b>A</b>) ABTS free radical scavenging abilities of ascorbic acid (0.1 mg/mL), MLP (0.1 mg/mL), and MLPHs (0.1 mg/mL). (<b>B</b>) DPPH scavenging activities of ascorbic acid (0.1 mg/mL), MLP (0.1 mg/mL), and MLPHs (0.1 mg/mL). (<b>C</b>) OH free radical scavenging activities of ascorbic acid (0.1 mg/mL), MLP (0.1 mg/mL), and MLPHs (0.1 mg/mL). (<b>D</b>) Reducing power of ascorbic acid (0.1 mg/mL), MLP (0.1 mg/mL), and MLPHs (0.1 mg/mL). Data are representative of three independent biological replicates. Data are means ± SDs. *: <span class="html-italic">p</span> ≤ 0.05; **: <span class="html-italic">p</span> ≤ 0.01; ***: <span class="html-italic">p</span> ≤ 0.001; and ****: <span class="html-italic">p</span> ≤ 0.0001.</p>
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<p>The isolation and characterization of bioactive mulberry peptides (BMPs) from mulberry leaf protein hydrolysates (MLPHs). (<b>A</b>) The separation chromatogram of MLPHs by DEAE Sepharose FF. F1-F6 are the six major isolates of MLPHs. (<b>B</b>) The DPPH radical scavenging activity of the F1-F6 fractions mentioned in (<b>A</b>). (<b>C</b>) The separation chromatogram of F3, F4, and F5 by Sephadex G-25. F3, F4, and F5 were all separated into 2 major components, which were exhibited as two peaks (F3-1, F3-2; F4-1, F4-2; F5-1, F5-2). (<b>D</b>) The DPPH radical scavenging activity of fractions F3-1, F3-2, F4-1, F4-2, F5-1, and F5-2 mentioned in (<b>C</b>). (<b>E</b>) The total ion chromatogram of F4-2. Data are representative of three independent biological replicates. Data are means ± SDs. *: <span class="html-italic">p</span> ≤ 0.05 and **: <span class="html-italic">p</span> ≤ 0.01.</p>
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<p>The isolation and characterization of bioactive mulberry peptides (BMPs) from mulberry leaf protein hydrolysates (MLPHs). (<b>A</b>) The separation chromatogram of MLPHs by DEAE Sepharose FF. F1-F6 are the six major isolates of MLPHs. (<b>B</b>) The DPPH radical scavenging activity of the F1-F6 fractions mentioned in (<b>A</b>). (<b>C</b>) The separation chromatogram of F3, F4, and F5 by Sephadex G-25. F3, F4, and F5 were all separated into 2 major components, which were exhibited as two peaks (F3-1, F3-2; F4-1, F4-2; F5-1, F5-2). (<b>D</b>) The DPPH radical scavenging activity of fractions F3-1, F3-2, F4-1, F4-2, F5-1, and F5-2 mentioned in (<b>C</b>). (<b>E</b>) The total ion chromatogram of F4-2. Data are representative of three independent biological replicates. Data are means ± SDs. *: <span class="html-italic">p</span> ≤ 0.05 and **: <span class="html-italic">p</span> ≤ 0.01.</p>
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<p>The screening of bioactive mulberry peptides (BMPs) with antioxidant activity by a cellular assay. (<b>A</b>) The effects of BMPs on the survival rate of HepG2 cells. HepG2 cells were treated with P1-P8 (25–200 μg/mL) for 24 h, and the survival rate was determined by CCK8. (<b>B</b>) The protective effects of P1-P8 against H<sub>2</sub>O<sub>2</sub>-induced damage in HepG2 cells. HepG2 cells were pretreated with P1-P8 (100 μg/mL) for 6 h and then treated with H<sub>2</sub>O<sub>2</sub> (160 μM) for 24 h. The cell survival rate was determined by CCK8. (<b>C</b>) Secondary mass spectra of P6 and P7. Data are representative of three independent biological replicates. Data are means ± SDs. *: <span class="html-italic">p</span> ≤ 0.05.</p>
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<p>The effects of P6 and P7 on ROS, CAT, and SOD levels in HepG2 cells. (<b>A</b>) HepG2 cells were pretreated with Trolox (0.1 mg/mL), P6 (5–10 μg/mL), or P7 (5–10 μg/mL) for 6 h and then treated with H<sub>2</sub>O<sub>2</sub> (160 μM) for 24 h. The cells were then stained with DCFH-DA and imaged by fluorescence microscopy (200×). (<b>B</b>) The fluorescence intensity in (<b>A</b>) was determined by a microplate reader. (<b>C</b>) The effects of P6 and P7 on CAT activity in HepG2 cells. Cells were treated as in (<b>A</b>). (<b>D</b>) The effects of P6 and P7 on SOD activity in HepG2 cells. Cells were treated as in (A). Data are representative of three independent biological replicates. Data are means ± SDs. **: <span class="html-italic">p</span> ≤ 0.01; ***: <span class="html-italic">p</span> ≤ 0.001; and ****: <span class="html-italic">p</span> ≤ 0.0001.</p>
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<p>The effects of P6 and P7 on chromatin damage in HepG2 cells. HepG2 cells were pretreated with Trolox (0.1 mg/mL), P6 (5–10 μg/mL), or P7 (5–10 μg/mL) for 6 h and then treated with H<sub>2</sub>O<sub>2</sub> (160 μM) for 24 h. The cells were then stained with Hoechst 33258 and imaged by fluorescence microscopy (200×).</p>
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<p>The protective effect of P6 and P7 against cell apoptosis. (<b>A</b>) HepG2 cells were pretreated with Trolox (0.1 mg/mL), P6 (5–10 μg/mL), and P7 (5–10 μg/mL) for 6 h and then treated with H<sub>2</sub>O<sub>2</sub> (160 μM) for 24 h. The cells were then stained with JC-1 and imaged by fluorescence microscopy (200×). (<b>B</b>) The fluorescence intensity in (<b>A</b>) was determined by a microplate reader. Cells were treated as in (<b>A</b>). (<b>C</b>) The detection of apoptosis by flow cytometry using the Annexin V–FITC assay. Cells were treated as in (<b>A</b>). (<b>D</b>) The effects of P6 and P7 on the apoptosis rate in HepG2 cells. Cells were treated as in (<b>A</b>). Data are representative of three independent biological replicates. Data are means ± SDs. *: <span class="html-italic">p</span> ≤ 0.05; **: <span class="html-italic">p</span> ≤ 0.01; ***: <span class="html-italic">p</span> ≤ 0.001.</p>
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