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Mitochondrial DNA sequence of the sub-unit 1 of cytochrome c oxidase (Cox1) and ribosomal DNA sequence of the first internal transcribed spacer (ITS-1) of Fasciola collected from mithun and cattle from Arunachal Pradesh, India were characterized at molecular level in order to identify the species of parasite. Based on sequence and phylogenetic analysis, the identity of the parasite was confirmed as Fasciola gigantica in mithun as well as from cattle. In order to know histopathological alteration in Fasciola infection, a histopathological study was performed on eight liver specimens, out of 110 animals studied during the period from 2010-2016. Histopathological examination of F.gigantica infected liver showed extensive fibrous connective tissue proliferation with necrosis of hepatocytes and infiltration of polymorphonuclear cell. There was evidence of migratory tracts of parasites with losing of normal lobular hepatic architecture. Bile duct proliferation was followed by congestion of portal vein with perivascular cuffing with surrounding degeneration of hepatocytes. The hepatocytes showed pyknosis with hyperplasia of bile duct. [ABSTRACT FROM AUTHOR]

The present study was conducted to assess the effect of the egg yolk powder in place of fresh egg yolk (EY) in the semen diluent on semen morphological attributes (SMAs) and mobility & velocity profiles measured by computer assisted sperm analyser (CASA) in cryopreserved mithun semen. A total of 25 ejaculates were collected through rectal massage method from five healthy mithun bulls (five ejaculates from each bull) and diluted with the standard tris citrate glycerol (TCG) extender and were splited into four equal aliquots, viz. Gr I, control (20% EY); Gr II, III and IV contained 5, 8 and 10% EY powder, respectively. SMAs, CASA parameters were evaluated following freezing-thawing of semen. Result revealed significant (P<0.05) improvement was observed in these SMAs and CASA parameters in Gr II and III as compared to Gr I and IV. It was concluded that inclusion of 5% or 8% EY powder in semen diluent holds a clear advantage and higher benefits over 20% fresh EY in cryopreservation of mithun semen. [ABSTRACT FROM AUTHOR]

The present study was designed to measure the effect of melatonin (MT) on apoptosis of sperm in mithun. Ejaculates (20) were collected from mithun bulls and were split into five equal aliquots, diluted with the TEYC extender. (Gr 1: semen without additives (control), Gr 2 to Gr 6: semen was diluted with 1, 2, 3, 4 and 5 mM of MT respectively). Apoptotic sperm percentage was estimated using commercially available apoptotic assessment kit at immediately after dilution, equilibration and post-thaw. Inclusion of MT into diluent resulted in a significant decrease in apoptotic sperm percentage at different stage of semen preservation as compared to untreated control group. Moreover, MT at 3 mM had significantly lower apoptotic sperm percentage than MT at 1 mM, 2 mM or 4 mM or 5 mM stored sperm at different stage. It was concluded that MT at 3 mM is suitable to minimize apoptosis in mithun sperm to pursuit future sperm preservation protocols. [ABSTRACT FROM AUTHOR]

It is now a well-accepted notion that each new experimental design requires proper evaluation of internal control genes (ICGs) for accurate normalization of expression data. In riverine buffaloes, till date no appropriate ICG has been reported for studying transcriptional response under any of the physiological stressful condition. The objective here was to test 16 well-known reference genes from different functional categories that could serve as suitable ICG during heat stress studies in buffalo mammary tissue. Briefly, the mammary explants were exposed to 45°C for 1 h and subsequently allowed to recover at 37°C for different time points (2-24 h). Three software programs, geNorm, Normfinder and BestKeeper, were used to measure gene transcript stability. RPL22 was excluded because of weak amplification and unacceptable PCR efficiency. Except GAPDH, all other genes showed expression stability within the acceptable range (<1.5). RPL4, B2M, RPS23 and EEF1A1 genes were found to be most stably expressed while GAPDH and ACTB showed least stability. The BestKeeper analysis identified high correlation for RPL4 ( r = 0.953) and EEF1A1 ( r = 0.914) with BestKeeper index. Based on the present findings, it could be suggested that geometric average of RPL4, B2M, RPS23 and EEF1A1 would provide accurate normalization to transcriptional data of buffalo mammary explant in response to heat stress. [ABSTRACT FROM AUTHOR]

The present study was undertaken to assess the effect of melatonin (MT) on sperm motility, viability, total sperm abnormality, acrosomal and plasma membrane integrity, DNA abnormality, antioxidant profiles such as superoxide dismutase (SOD), catalase (CAT), glutathione (GSH) and total antioxidant capacity (TAC), enzymatic profiles such as aspartate amino transaminase (AST), alanine amino transaminase (ALT), and biochemical profiles such as malonaldehyde (MDA) production and cholesterol efflux. Total numbers of 30 ejaculates were collected twice a week from eight mithun bulls and semen was split into five equal aliquots, diluted with the TEYC extender. Group 1 has semen without additives (control) and group 2 to group 5 have semen that was diluted with 1mM, 2mM, 3mM, and 4mMof melatonin, respectively. These seminal parameters, antioxidant, enzymatic, and biochemical profiles were assessed at 5°C for 0, 6, 12, 24, and 30 h of incubation. Inclusion of melatonin into diluent resulted in significant (p < 0.05) decrease in percentages of dead spermatozoa, abnormal spermatozoa, and acrosomal abnormalities at different hours of storage periods as compared with control group. Additionally, melatonin at 3mMhas significant improvement in quality of mithun semen than melatonin at 1mM, 2mMor 4mMstored in in vitro for up to 30 h. It was concluded that the possible protective effects of melatonin on sperm parameters are it prevents MDA production and preserve the antioxidants and intracellular enzymes during preservation. [ABSTRACT FROM AUTHOR]

The aim of the present study was to evaluate the effect of feeding green fodder, rice straw and concentrate-based total mixed rations (TMR) on dry matter (DM) intake (DMI), nutrient utilization, rumen fermentation patterns and body weight (BW) gain (BWG) in mithun ( Bos frontalis) calves. In a randomized block design, male mithun calves ( n = 18, 8–10 months of age, 121 ± 2 kg BW) were randomly divided into three experimental equal groups (six animals in each group) and fed isonitrogenous TMRs ad libitum for 120 days . The TMR1 contained 30% Napier grass and 30% rice straw, TMR2 contained 60% rice straw and TMR3 contained 30% tree leaves ( Lagerstroemia speciosa) and 30% rice straw (DM basis). All the TMRs contained 40% concentrate mixture (DM basis). The results indicated that the BWG, DMI and feed conversion efficiency were significantly (p < 0.01) increased with the inclusion of green fodder in TMRs. The apparent digestibility of DM, crude protein, ether extract, crude fibre and nitrogen free extract were also improved significantly (p < 0.01) with the inclusion of green fodder in TMRs. The higher concentration of total nitrogen and total volatile fatty acid in rumen liquor, but low ruminal pH were evident in animals fed green fodder supplemented TMRs. An increased (p < 0.01) molar proportion of acetic acid was evident in animals fed rice straw-based TMR. In contrast, the molar proportion of propionic and butyric acids were significantly (p < 0.01) higher in animals fed green fodder supplemented TMRs. On the basis of higher DMI and higher daily BWG, it is concluded that Napier grass and L. speciosa tree leaves may be incorporated upto 30% (DM basis) in TMR of growing mithuns for feeding in complete confinement system. [ABSTRACT FROM AUTHOR]

The present study was designed to develop and validate a simple and sensitive radioimmunoassay (RIA) procedure to estimate FSH in mithun (Bos frontalis) plasma. The assay was carried out in 100 µL of mithun plasma. The bovine FSH standards (10 to 5000 pg/100 µL/tube) in hormone-free plasma were used in the assay. The sensitivity of the assay was 20 pg/100 µL/tube, which corresponded to 0.20 ng/mL plasma. The 50% relative binding sensitivity of the assay was 80 pg/100 µL/tube, which corresponded to 0.80 ng/mL plasma. The intra- and inter-assay coefficients of variation were 4.6% and 12.4%, respectively. The biological validation of the assay was carried out in plasma samples that were collected during different stages of the estrous cycle. In the entire estrous cycle, plasma FSH concentration (p<0.01) attained two peaks (on day 3 to 4 before estrus 5.1±0.3 ng/mL and on the day of estrus 6.9±0.2 ng/mL). FSH concentration remained at basal level (1.3±0.1 to 1.6±0.2 ng/mL) during day 4 to 16 of the estrous cycle. The concentration of plasma FSH was found to be significantly (p<0.05) higher (4.9±0.3 to 6.8±0.5 ng/mL) until 48 h following the estrus onset. In conclusion, the RIA procedure that was developed in the current study is sufficiently reliable and sensitive to estimate different physiological levels of FSH in mithun plasma. [ABSTRACT FROM AUTHOR]

Contents The objective of the present study was to establish the changes in plasma concentrations of LH, FSH, estradiol 17- β (E2) and progesterone (P4), as well as to understand their temporal relationships during oestrus in mithun ( Bos frontalis). The experiment was conducted on 11 mithuns during third or fourth postpartum oestrous cycle. Since oestrus onset the jugular vein blood samples were collected every 2 h for 72 and 96 h, respectively from the animals without and with standing heat. The LH, FSH, E2 and P4 concentrations were estimated in plasma. The P4 concentration was fluctuated throughout the oestrus period and the average P4 concentration was found significantly (p < 0.05) lower on the day of oestrus onset. The multiple rises in LH and FSH concentrations above the basal level in spike like fashion were observed throughout the oestrus period irrespective of the occurrence of standing heat. A significant (p < 0.01) gradual increase in the average daily E2 concentration was observed till day 2 following oestrus onset irrespective of the occurrence of standing heat. A significant (p < 0.05) simultaneous increase in LH, FSH and E2 concentrations and a transient increase in P4 concentration at approximately the time of standing heat onset were observed. During investigation a definite temporal coupling between LH and FSH rises was absent throughout the oestrus period. The results suggest that (1) the multiple short-duration low-amplitude LH and FSH surges during oestrus may be crucial for the final maturation of ovulatory follicle and subsequent ovulation in mithun; (2) a differential mechanism for controlling LH and FSH secretions probably exists in mithun. [ABSTRACT FROM AUTHOR]

In domestic species, spontaneous erection and penile movements or manipulation occur in an awake state, the behaviour is commonly known as masturbation. Six mithun bulls were selected, of which, three from Mizoram and three from Manipur strain. These strains were observed over a period of 12 months. This behaviuor were observed from 1800 to 800 hrs and especially in winter and spring season. The parameters studied were duration of erection, frequency of movements of penis, time of sexual erection and masturbation. Seminal parameters such as volume, mass activity, progressive motility, live sperm, concentration, abnormality, acrosomal integrity and HOST were studied, which were collected by massage method. Other parameters such as success rate, protrusion time, length of penis and ejaculation time were also studied. The seminal parameters were showed that there was no significant difference between the affected and unaffected animal. But the unaffected animals were non-significantly higher in seminal parameter than affected animals. [ABSTRACT FROM AUTHOR]

The article presents a study which discusses the helmintic infections on mithuns. A collection of fresh faecal samples from 93 mithuns have been observed from different areas in the Phek district of Nagaland, India including Porba, Thevopisu, and institute farms in Porba. Results indicate that 81.7% among the 76 animals have been positive for various helmintic infections wherein the most common is Trichostrongylus, Cooperia, and Haemonchus.

The handmade cloning technique has been a relatively recent addition in the field of nuclear transfer. In the present study, attempts were made to efficiently derive stem cells from handmade cloned (HMC) embryos in goat using adult fibroblast cells, embryonic stem (ES) cells, and lymphocytes as donor cells, and to characterise the derived putative nuclear transfer ES (ntES) cells for their stemness. Efficiency of the donor cells for nuclear transfer was also compared, and an overall cleavage and morula formation rates of 62.44±3.9% and 35.30±3.86%, 75.45±3.92% and 45.84±3.86%, and 56.38±3.92% and 29.09±3.86% were obtained from adult fibroblasts, ES cells, and lymphocytes, respectively. A significant difference was found between ES cells and the other 2 donor cells in terms of cleavage and morula formation. However, no such difference existed between fibroblasts and lymphocyte donor cells. Stem cell colonies were successfully derived from HMC embryos obtained from all 3 different donor cells. The rate of primary colony formation was 61.66±4.62% for fibroblast-donor-cell-derived embryos. This rate was 59.91±4.62% for ES-donor-cell-derived embryos and 62.49±4.62% for lymphocyte-donor-cell-derived embryos. The putative ntES colonies were positively characterised for TRA-1-60, TRA-1-81, SSEA-1, SSEA-4, OCT-4, SOX-2, and Nanogby immunocytochemistry and RT-PCR. Results indicated that ES cells had better efficiency as donor cells in cloned embryo production than did adult fibroblasts and lymphocytes. The finding also suggested that terminally differentiated cell-like lymphocytes can also be reprogrammed. Moreover, there was no difference between the different donor-cell-derived HMC embryos in terms of ntES cell derivation. The study has established an efficient protocol for putative ntES cell derivation from HMC embryos. This could be of substantial significance because patient-specific ntES cells have proven therapeutic significance. The authors acknowledge N.D.R.I for the financial and infrastructural assistance. [ABSTRACT FROM AUTHOR]

The article presents a case report of tick infestation in Bos frantalis or mithun animals at Nagaland, India. Ticks were examined from the external body parts of 102 mithun animals reared at Jharnapani and 26 animals of Porba farms of the Institute and 48 animals free range animals from Porba village. From the examination made, the Ixodid tick of the species Boophilus microplus was identified as the only ectoparasite. The inner side of pinna and legs are the common predilection sites of ticks.

Abstract: The objective of the present study was to investigate the possibility of preserving mithun (Bos frontalis) spermatozoa at refrigeration temperature using tris–egg yolk diluent. Semen samples were collected from four adult mithun bulls through rectal massage method. Good quality semen samples (n =30) were preserved at 4°C using tris–egg yolk diluent for 72h. Progressive motility, live spermatozoa count and morphological abnormalities were evaluated every 12h until 72h of preservation. The colour, consistency and mass activity of fresh semen samples were found to be creamy white, medium and 3+ to 4+ (5+ scale), respectively. The average (mean±S.E.) volume (ml), pH and spermatozoa concentration (106 ml−1) of fresh semen samples were found to be 0.6±0.01, 6.8±0.03 and 425±48, respectively. Progressive motility and live spermatozoa count were found to be less than 30% (P <0.01) after 48h of storage. Head (P <0.05), midpiece (P <0.05), tail (P <0.01) and total (P <0.01) abnormalities were found to be increased significantly over the time of storage. It was observed that progressive motility and live spermatozoa count remained above 30% and 40%, respectively, until 36h of storage. Simultaneously the percentage of morphologically abnormal spermatozoa was found to be significantly low until 36h of storage. The results indicate that it is possible to preserve mithun spermatozoa at refrigeration temperature in tris–egg yolk diluent, which can be further used for artificial insemination within 36h of storage. [Copyright &y& Elsevier]

Mithun (Bos frontalis), a domestically raised herbivore, holds significant economic importance for the farming community of Northeast India. This study aimed to elucidate the genetic parameters governing Mithun body weight traits across different ages using data from the sole organized semi-intensive Mithun farm in India. Information was gathered from 110 Mithuns born over a period spanning from 2011 to 2022. Body weight taken at week 1 (W1), 1-month (M1), 3-months (M3), 6-months (M6), 9-months (M9), 12-months (M12), 30-months (M30) and 45-months (M45) were considered for the study. The genetic parameters estimation employed the BLUPF90 suite of programs, incorporating univariate Gibbs sampler animal model with fixed effects; season and period of birth, and sex of the animal. Variance and covariance components, including direct additive genetic effects, were estimated. Heritability estimates for the eight body weight traits ranged from 0.47 ± 0.0050 to 0.50 ± 0.0043, indicating varying genetic influence across growth stages. Results revealed that Mithun herd has a substantial genetic variability for growth traits and therefore there is ample scope to select for a better growth rate. Here, we conclude that Month 12 (M12) and Month 9 (M9) body weights exhibit higher heritability, indicating potential for genetic improvement through selective breeding., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

The Flinders Technology Associates (FTA) card, a cotton-based cellulose membrane impregnated with a chaotropic agent, effectively inactivates infectious microorganisms, lyses cellular material, and fixes nucleic acid. The aim of this study is to assess the stability and detection limit of various RNA viruses, especially the avian influenza virus (AIV), Newcastle disease virus (NDV), and African horse sickness virus (AHSV), on the FTA card, which could significantly impact virus storage and transport practices. To achieve this, each virus dilution was inoculated onto an FTA card and stored at room temperature in plastic bags for durations ranging from 1 week to 6 months. Following storage, the target genome was detected using conventional reverse transcription polymerase chain reaction. The present study demonstrated that the detection limit of AIV ranged from 1.17 to 6.17 EID 50 values over durations ranging from 1 week to 5 months, while for NDV, it ranged from 2.83 to 5.83 ELD 50 over the same duration. Additionally, the detection limit of AHSV was determined as 4.01 PFU for both 1 and 2 weeks, respectively. Based on the demonstrated effectiveness, stability, and safety implications observed in the study, FTA cards are recommended for virus storage and transport, thus facilitating the molecular detection and identification of RNA viral pathogens.

Lumpy skin disease (LSD) is a disease of cattle that is also known to cause mild infection in buffaloes. To date, there have been no reports of LSD in mithun (Bos frontalis), a bovine species distributed in Northeast India, Bangladesh, Myanmar, and parts of China. In the present study, the presence of typical clinical signs, virus isolation, PCR amplification, sequence analysis, and the demonstration of antibodies in serum by indirect enzyme-linked immunosorbent assay and serum neutralization test, confirmed the occurrence of LSD in mithun for the first time in India. Phylogenetic analysis based on the full-length RPO30 and P32 genes of LSD virus from mithun and cattle revealed 100% sequence identity, indicating circulation of the same strain in both species in India and the possibility of spillover between species., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature.)

The present study aimed to determine carcass characteristics, meat quality, nutrient profiles, and sensory characteristics of Mithun meat. Sixteen Mithun were selected and divided into four groups, MM-4 (male; n = 4; <4 years of age), MM-47 (male; n = 4; 4-7 years of age), MF-4 (female; n = 4; <4 years of age), MF-47 (female; n = 4; 4-7 years of age). Carcass characteristics showed that adult males (MM-47) have significantly higher (P < 0.05) live weight, carcass weight, and meat-to-bone ratio. Fat (%) was significantly higher (P < 0.05), and deboned meat (%) was lower in MF-4 and MF-47, while marginal differences were observed in bone (%), dressing percentage, and offal yield between groups. Visible marbling increased with age and varied from "slight" to "small" in all groups. Nutrient profiling revealed a significantly higher (P < 0.05) fat percentage and cholesterol in MF-4 and MF-47. Fatty acid profile, amino acid profile, water-soluble vitamins, and minerals content did not differ between groups. However, lysine and leucine (essential amino acids) and glutamic acid and aspartic acid (nonessential amino acids) were most abundant. Effect of age was significant (P < 0.05) on juiciness, tenderness, and connective tissue residue scores. In conclusion, results indicate mithun meat is nutrient-rich regardless of the animal's age or sex., (© 2024 Japanese Society of Animal Science.)

Seeking a new drug has become a significant milestone in drug discovery. However, it might not be immediately used in urgent situations or during a pandemic. Acute Respiratory Distress Syndrome (ARDS) contributes to mild-to-severe symptoms in patients due to cytokine storms, leading to morbidity and mortality. Hypertension is recognized as an independent risk factor for the severity of ARDS regarding to both ACE Inhibitors (ACEIs) and Angiotensin Receptor Blockers (ARBs) treatment, although the precise mechanism remains unclear. In this study, murine macrophage cell lines (RAW264.7) and alveolar epithelial type II-like cell lines (A549) were utilized to investigate the effect of Losartan (LOS). LOS attenuated nitric oxide production in a dose-dependent manner and collectively reduced intracellular reactive oxygen species (ROS) compared to Diclofenac under LPS-stimulation conditions. For ADRS-mimicking conditions, LPS-induced inflammatory A549 cells were performed to monitor the effect of LOS. The results showed that LOS exhibited a protective effect by increasing cell viability and decreasing intracellular ROS levels. Notably, a high dose of LOS increased intracellular ROS levels. Moreover, LOS treatment downregulated NF-kappaB activation and AT1R at the protein level. Correspondingly, proinflammatory mediator cytokines (TNF-alpha and IL-8) were downregulated, but not IL-6, during LOS treatment. Hence, LOS may provide substantial benefits to ARDS patients by modulating proinflammatory cytokine production through AT1R downregulation and NF-kappaB inactivation. The mechanistic insight into LOS's anti-inflammatory effect holds promise for reducing mortality rates among ARDS patients., (© The Author(s).)

Oriental theileriosis caused by Theileria orientalis, previously considered a benign disease, is posing a significant threat to the livestock industry across the globe. To elucidate the prevalence of Theileria orientalis in ticks and their host, the Mithun, a comprehensive study was undertaken in the two northeastern states of India, viz. Nagaland and Arunachal Pradesh. A total of 340 of Rhipicephalus microplus ticks and 25 Ambylomma sp. ticks were screened for the presence of Theileria orientalis through PCR. Among the R. microplus ticks examined, 25 of them tested positive for T. orientalis infection whereas none of the Amblyomma ticks was positive. Additionally, a total of 275 blood samples were collected from Mithun from Arunachal and Nagaland and 31 animals were found to be positive for T. orientalis infection. Notably, six positive cases were identified in Porba (Phek district), six in Tening, and one in Bamsiakilwa village (Peren district) of Nagaland. Moreover, out of the 41 animals examined at Medziphema farms, Nagaland, 18 were found to be positive for T. orientalis infection. Moreover, the phylogenetic investigation has unveiled the presence of the highly pathogenic Type 2 (Ikeda) T. orientalis genotype in Mithun, supported by a strong bootstrap value of 100%. This study marks the initial documentation of oriental theileriosis in mithun. It underscores the need for vigilant monitoring and active surveillance of mithun populations in the northeastern states of India. Timely treatment of infected animals is imperative to avert economic losses for the farmers., (© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Background and Aim: African horse sickness (AHS) has become a newly emerging disease after an outbreak in northeastern Thailand in March 2020. Mass vaccination in horses with live-attenuated AHS virus (AHSV) vaccine is essential for AHS control and prevention. This study aimed to monitor the longitudinal humoral immune response before and after a single vaccination using a live-attenuated vaccine against AHS in stallions, mares, and pregnant mares, including maternal immunity in foals born from pregnant mares during the outbreak in Thailand., Materials and Methods: A total of 13 stallions and 23 non-pregnant and 21 pregnant mares were vaccinated with live-attenuated AHSV vaccines. Serum samples from selected horses were collected on the day of vaccination and 1, 6, 8, 9, 10, and 12-months post-vaccination. Furthermore, seven serum samples of foals born from vaccinated pregnant mares were collected on parturition date and 1, 3, and 6-months old. The antibody titer against AHS in all collected serum samples was evaluated using a commercial enzyme-linked immunosorbent assay kit. All data were analyzed for mean and standard deviation for each group of samples using a spreadsheet program. Antibody titers between times were analyzed using a one-way analysis of variance as repeated measurement, and antibody titers between horse groups were analyzed using a general linear model for statistically significant differences when p < 0.05., Results: In stallion and non-pregnant mare groups, there were no statistically significant differences in antibody titers in all 6 time periods after vaccination. The antibody titer in the pregnant mare group showed a non-statistically significant difference between each gestation stage, except at 8 months post-vaccination. Furthermore, increasing antibody titers on days 1 and 3 after receiving colostrum in foals indicate the major role of transcolostral antibody transfer for AHS., Conclusion: This study demonstrated that a single AHS vaccination using a live-attenuated vaccine could stimulate high antibody titers sufficient for AHS control and prevention during the outbreak in Thailand. Similarly, the antibody response of vaccinated horses of both genders, including various stages of pregnant mares, was statistically not different., Competing Interests: The authors declare that they have no competing interests., (Copyright: © Kunanusont, et al.)

Background: Abnormal blood lipid levels in the general population and adverse reproductive events among women have been associated with cognitive impairment (CI). However, their relationship has not been extensively studied in community settings. Hence, this study aims to explore the association of CI with blood lipid levels in both sexes and reproductive events/trajectory among women., Methods: A cross-sectional study was conducted among a North Indian rural population. A total of 808 adults were recruited through door-to-door household survey. Data on socio-demographic variables, reproductive profile of women, and cognitive impairment status were collected. Fasting blood sample was collected to estimate serum lipid profile. Multivariate logistic regression was performed to test for association., Results: The study demonstrated a lack of association between lipid profile and cognitive impairment among males. Surprisingly, low HDL-C among females was found to be protective against moderate/severe cognitive impairment (value of p  = 0.049). Further, menopausal women and those having five or higher live births were found to be at higher risk of CI than pre-menopausal women and those with 1-2 live births, respectively., Conclusion: The present study hints toward a gender-specific association of blood lipid levels with CI. Further, higher live births and menopause appear to be important risk factors for CI among women., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Khate, Chaudhary, Longkumer, Saraswathy and Devi.)

Background and Aim: The immune responses of animals infected with African horse sickness (AHS) virus are determined by enzyme-linked immunosorbent assay (ELISA), complement fixation, and virus neutralization test. During the outbreaks of AHS in Thailand, the immune response after vaccination has been monitored using commercial test kits such as blocking ELISA, which are expensive imported products unavailable commercially in Thailand. This study aimed to assess the sensitivity and specificity of anti-AHS virus antibodies using dot blotting based on monovalent and polyvalent strains of live attenuated AHS vaccine., Materials and Methods: A total of 186 horse sera, namely, 93 AHS-unvaccinated samples and 93 AHS-vaccinated samples, were used in this study. All sera underwent antibodies detection using commercial blocking ELISA and in-house dot blotting based on monovalent and polyvalent strains of live attenuated AHS vaccine. The numbers of true positive, false positive, true negative, and false negative results in the dot blotting were compared with those in blocking ELISA and the sensitivity and specificity of dot blotting were assessed., Results: For the monovalent antigen, there were 78, 19, 74, and 15 true positive, false positive, true negative, and false negative results, respectively, while for the polyvalent antigen, the corresponding numbers were 84, 34, 58, and 9. Meanwhile, the diagnostic sensitivity and specificity for monovalent antigen were 83.87% and 79.57%, respectively, but 90.32% and 62.37% for polyvalent antigen., Conclusion: Dot blotting for AHS antibodies detection using vaccine antigen showed high sensitivity and rather a high specificity compared with the findings with the commercial ELISA test kit. In countries where commercial ELISA test kits are not available and when the size of a serum sample is small, dot blotting could become a good alternative test given its advantages, including its simplicity, rapidity, and convenience. To the best of our knowledge, these findings are the first report on the use of dot blotting for detecting AHS antibodies in horses. In conclusion, monovalent antigen-based dot blotting could be used as a reliable alternative serodiagnostic test for monitoring AHS humoral immune response, especially in vaccinated horses., Competing Interests: The authors declare that they have no competing interests., (Copyright: © Taesuji, et al.)

Background and Aim: The flinders technology associates (FTA) card is a cotton-based cellulose membrane impregnated with a chaotropic agent that inactivates infectious microorganisms, lyses cellular material, and fixes DNA and/or RNA within the fiber matrix. However, little is known about the effectiveness of these cards for detecting RNA viruses in animals. This study aimed to evaluate the sensitivity of RNA virus detection using conventional reverse-transcription polymerase chain reaction (RT-PCR) on FTA cards., Materials and Methods: A highly virulent Newcastle disease virus (NDV) and an avian influenza virus (AIV) with low pathogenicity were propagated using chicken embryonic eggs. Three days after inoculation, the allantoic fluid was harvested, stored at -80°C, and the stock virus was tested for virus titration. African horse sickness virus (AHSV) was obtained from a live attenuated vaccine that was dissolved and stored at -80°C. For sample preparation, each stock virus was 10-fold serially diluted and each dilution was inoculated onto an FTA card, followed by drying in a Class II safety cabinet. Both the stock virus and infected FTA card were genomically isolated using an extraction kit, FTA purification kit, and extraction kit with Tris-EDTA (TE) buffer. The target genome was then detected by one-step RT-PCR for NDV and AIV, and two-step RT-PCR for African horse sickness, including gel electrophoresis for the detection of specific nucleic acids., Results: The detection limit of stock AIV was compared on FTA cards, using the FTA purification kit, and with TE buffer with an extraction kit. The corresponding results were 1.47, 1.17, and 2.18 log 10 EID 50 , respectively, while for NDV the results were 4.13, 4.83, and 4.84 log 10 ELD 50 . Finally, detection limit of stock AHSV and AHSV on the FTA card extracted using TE buffer with an extraction kit were 4.30 and 4.01 log 10 plaque-forming units, respectively., Conclusion: This study demonstrated that the detection limit or sensitivity of all tested RNA viruses on FTA cards did not differ when compared with those of the stock virus and in both methods for RNA isolation on FTA cards. These cards are suitable for collecting and transporting samples infected with RNA viruses, particularly AIV, NDV, and AHSV. Flinders technology associates cards also provide hazard-free samples, a reliable source of RNA for molecular characterization, and sufficient quantity for diagnostic applications based on nucleic acid-based detection., Competing Interests: The authors declare that they have no competing interests., (Copyright: © Rattanamas, et al.)

The study aimed to evaluate follicular dynamics and concentrations of estradiol-17β (E2), progesterone (P4), follicle stimulating hormone (FSH) and luteinizing hormone (LH) during the oestrous cycle and to determine ovulation time in Mithun cows. Ovaries of experimental cows (n = 7) were examined daily by transrectal-ultrasonography for three consecutive oestrous cycles (n = 21). The characteristics of follicular waves, dominant follicle, largest subordinate follicle and corpus luteum and ovulation time were evaluated. The plasma samples were analysed throughout the interovulatory interval to determine the differences in the hormonal profiles (E2, P4, FSH and LH) between different follicular wave cycles. Out of eighteen oestrous cycles analysed, three-wave follicular cycles were maximum (n = 12: 66.66%) followed by two (n = 4: 22.22%) and four waves (n = 2: 11.11%). The two and three waves were statistically compared, and no significant (p > .05) differences were observed in day of wave emergence, number of follicles (≥3 mm) recruited, maximum diameter of the ovulatory dominant follicle, growth rates of ovulatory and anovulatory dominant follicles and maximum diameter of corpus luteum. The diameter of dominant follicles was significantly (p < .05) greater than subordinate follicles in both ovulatory and anovulatory waves. No significant differences were observed in peak concentrations of estradiol-17β and follicle stimulating hormone between ovulatory and anovulatory waves in all wave cycles. A preovulatory luteinizing hormone surge was observed a day before ovulation in all wave cycles. Progesterone concentrations were lower than 0.5 ng/ml during oestrus and increased sharply to the maximum levels of ≥3.8 ng/ml in all wave cycles. Ovulation time (mean ± SEM), irrespective of follicular waves was 10.5 ± 0.64 h after the end of oestrus. It was concluded that Mithun cows have a preponderance of three follicular waves with little difference between the two- and three-follicular waves and ovulation occurred 10.5 h after the end of oestrus., (© 2022 Wiley-VCH GmbH.)

Efficacy of African swine fever virus (ASFV) inactivation using five commercially supply compound disinfectants was evaluated under various condition. Virucidal efficacy demonstrated that products A and E could inactivate at 1:800 within 1 min for both temperatures, while products B, C and D inactivated at 1:400. However, product D could inactivate at 1:800 when the exposure time was extended to 30 min and effected only 20°C. In addition, the cytotoxicity demonstrated that products A, B, C, D and E did not significantly affect to cell at 1:51,200, 1:12,800, 1:12,800, 1:25,600 and 1:12,800 dilution, respectively. In conclusion, these disinfectants could inactivate ASFV, however, the application of these products should be performed under safety precautions to prevent cytotoxicity in humans and animals.

Background and Aim: The selection and proper application of disinfectants are crucial to the prevention of many diseases, so disinfectants must be evaluated before being used for the prevention of African swine fever (ASF). Three disinfectant products belonging to the group of potassium hydrogen peroxymonosulfates, product A and product B, and a quaternary ammonium compound called product C, were examined in vitro for host cell cytotoxicity and the efficacy of ASF virus inactivation. The study parameters included various concentrations, exposure times, temperatures, and degrees of cytotoxicity., Materials and Methods: Three disinfectant products were evaluated for cytotoxicity using primary porcine alveolar macrophage (PAM) cells at dilutions from 1:200 to 1:51,200. Disinfectants in concentrations of 1:200, 1:400, and 1:800 were prepared, the pH and the virucidal activity were tested. An equal volume of each dilution was mixed with the ASF virus and incubated at room temperature (20°C) or on ice (4°C) for 1 min, 5 min, or 30 min. Hemadsorption (HAD) or rosette formation was observed using an inverted microscope for 5 days after inoculation, and the virus titer was calculated as HAD 50 /mL. Each treatment and virus control were tested in triplicate, and the titers were reported as means and standard deviations. The reduction factor was used to measure inactivation., Results: Products A, B, and C at 1:400, 1:800, and 1:25,600 of dilution, respectively, did not show significant cytotoxic effects on PAM cells. Products A and B could inactivate ASF virus at 1:200 dilution within 5 min after exposure at 4°C. However, at 20°C, the exposure time had to be extended to 30 min to inactivate the virus. Product C could inactivate the virus at 1:400 dilution within 5 min under both temperature conditions, whereas at 1:800 dilution, the exposure time had to be extended to 30 min to completely inactivate the virus at 20°C., Conclusion: All disinfectants could inactivate ASF virus in various concentrations, under appropriate exposure times and reaction temperatures, and there was no evidence of host cell cytotoxicity. For the control of ASF in pig farms, the appropriate concentration, ambient temperature, and contact time of these disinfectants should be taken into account., (Copyright: © Sovijit, et al.)

A massive bovine, Bos frontalis, also known as Mithun or Gayal, found at higher altitude is very promising meat and milk animal. For candidate gene and marker discovery, RNA-seq data was generated from longissimus dorsi muscle tissues with Illumina-HiSeq. Such markers can be used in future for genetic gain of traits like feed conversion efficiency (FCE) and average daily gain (ADG). Analysis revealed 297differentially expressed genes (DEGs) having 173 up and 124 down-regulated unigenes. Extensive conservation was found in genic region while comparing with Bos taurus. Analysis revealed 57 pathways having 112 enzymes, 72 transcriptional factors and cofactors, 212 miRNAs regulating 71 DEGs, 25,855 SSRs, mithun-specific 104,822 variants and 7288 indels, gene regulatory network (GRN) having 24 hub-genes and transcriptional factors regulating cell proliferation, immune tolerance and myogenesis. This is first report of muscle transcriptome depicting candidate genes with GRN controlling FCE and ADG. Reported putative molecular markers, candidate genes and hub proteins can be valuable genomic resources for association studies in genetic improvement programme., (Copyright © 2019. Published by Elsevier Inc.)

Background: Mithun (Bos frontalis), also called gayal, is an endangered bovine species, under the tribe bovini with 2n = 58 XX chromosome complements and reared under the tropical rain forests region of India, China, Myanmar, Bhutan and Bangladesh. However, the origin of this species is still disputed and information on its genomic architecture is scanty so far. We trust that availability of its whole genome sequence data and assembly will greatly solve this problem and help to generate many information including phylogenetic status of mithun. Recently, the first genome assembly of gayal, mithun of Chinese origin, was published. However, an improved reference genome assembly would still benefit in understanding genetic variation in mithun populations reared under diverse geographical locations and for building a superior consensus assembly. We, therefore, performed deep sequencing of the genome of an adult female mithun from India, assembled and annotated its genome and performed extensive bioinformatic analyses to produce a superior de novo genome assembly of mithun., Results: We generated ≈300 Gigabyte (Gb) raw reads from whole-genome deep sequencing platforms and assembled the sequence data using a hybrid assembly strategy to create a high quality de novo assembly of mithun with 96% recovered as per BUSCO analysis. The final genome assembly has a total length of 3.0 Gb, contains 5,015 scaffolds with an N50 value of 1 Mb. Repeat sequences constitute around 43.66% of the assembly. The genomic alignments between mithun to cattle showed that their genomes, as expected, are highly conserved. Gene annotation identified 28,044 protein-coding genes presented in mithun genome. The gene orthologous groups of mithun showed a high degree of similarity in comparison with other species, while fewer mithun specific coding sequences were found compared to those in cattle., Conclusion: Here we presented the first de novo draft genome assembly of Indian mithun having better coverage, less fragmented, better annotated, and constitutes a reasonably complete assembly compared to the previously published gayal genome. This comprehensive assembly unravelled the genomic architecture of mithun to a great extent and will provide a reference genome assembly to research community to elucidate the evolutionary history of mithun across its distinct geographical locations.

The current study aimed at genomic characterization and improved understanding of genetic diversity of two Indian mithun populations (both farm, 48 animals and field, 24 animals) using genome wide genotype data generated with Illumina BovineHD BeadChip. Eight additional populations of taurine cattle (Holstein and NDama), indicine cattle (Gir) and other evolutionarily closely related species (Bali cattle, Yak, Bison, Gaur and wild buffalo) were also included in this analysis (N = 137) for comparative purposes. Our results show that the genetic background of mithun populations was uniform with few possible signs of indicine admixture. In general, observed and expected heterozygosities were quite similar in these two populations. We also observed increased frequencies of small-sized runs of homozygosity (ROH) in the farm population compared to field mithuns. On the other hand, longer ROH were more frequent in field mithuns, which suggests recent founder effects and subsequent genetic drift due to close breeding in farmer herds. This represents the first study providing genetic evidence about the population structure and genomic diversity of Indian mithun. The information generated will be utilized for devising suitable breeding and conservation programme for mithun, an endangered bovine species in India.

Eleven reference genes from different functional categories were assessed for their stable expression pattern in heat stressed PBMCs across Indian buffalo and cattle using GeNorm, NormFinder and BestKeeper algorithms. As the first report, we suggest B2M, RPS9 and RPS15a as suitable reference genes for accurate normalization of PBMC transcript data while the ACTB gene is not recommended., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

The main purpose of the experiment was to compare the efficiency of three cell types, namely adult fibroblast, putative embryonic stem (ES) cell, and lymphocyte, as donor cells for somatic cell nuclear transfer by handmade cloning in goats. The outcome clearly shows that putative embryonic stem cells, with a cleavage and blastocyst production rate of 74.69% ± 3.92 and 39.75% ± 3.86, respectively, performs better in comparison to adult fibroblast cell and lymphocyte. Between adult fibroblast cell and lymphocyte no statistically significant difference exists at P < 0.05. An overall cleavage and blastocyst formation rate of 67.41% ± 3.92 and 26.96% ± 3.86 was obtained using adult fibroblast donor cells. The study establishes beyond doubt the reprogrammability of lymphocyte by handmade cloning (HMC) protocol with a cleavage and blastocyst production rate of 56.47% ± 3.92 and 24.70% ± 3.86, respectively. PCR analysis of highly polymorphic 286 bp fragment of MHC II DRB genes of cloned embryos and three donor cells were performed to verify the cloned embryos. The amplified PCR products were subjected to SSCP to confirm their genetic identity. The karyotyping of the cloned embryos showed normal chromosomal status as expected in goat. Significantly, in the second stage of the experiment, the produced cloned embryos were successfully used to derive ntES-like cells. The rate of primary colony formation rate was 62.50% ± 4.62 for fibroblast donor cell derived embryos. The same was 60.60% ± 4.62 for putative ES donor cell derived embryos and 66.66% ± 4.62 for lymphocyte donor cell derived embryos, respectively. The putative ntES colonies were positively characterized for alkaline phosphatase, Oct-4, TRA-1-60, TRA-1-81, Sox-2, and Nanog by Immunocytochemistry and Reverse Transcription PCR. To further validate the stem ness, the produced putative ntES colonies were differentiated to embryoid bodies. Immunocytochemistry revealed that embryoid bodies expressed NESTIN specific for ectodermal lineage; GATA-4 for endodermal lineage and smooth muscle actin-I, and troponin-I specific for mesodermal lineage. The study has established an efficient protocol for putative ntES cell derivation from HMC embryos. It could be of substantial significance as patient specific ntES cells have proven therapeutic significance., (Copyright © 2011 Elsevier Inc. All rights reserved.)

The objective of the study was to evaluate the effects of genetic and non-genetic factors on production of breeding bulls and semen quality parameters in Karan Fries crossbred male by fitting least squares analysis. Genetically, the animals were divided into three subclasses. The non-genetic factors were season of birth, period of birth, and age group with three subclasses each for season of birth and period of birth. Age group was classified into four subclasses. The traits generated in the study were number of males reaching semen donation stage (AFSC) and first freezing (AFSF), age at last semen collection (ALSC) and last freezing (ALSF), age at disposal (AD), and lifetime semen production traits (up to 1 year after first freezing). The effect of period of birth was significant for AFSC, AFSF, ALSC, and AD. It was also significant for total ejaculates produced in a year. The age group had significant effect on AFSF. Effect of genetic group was significant for freezable ejaculates produced in a year, for frozen semen doses produced in a year, and for number of ejaculates cryoprocessed in a year. Season had no statistically significant effect on any of the traits studied. The influence of period revealed that the most of the traits of breeding bulls improved after intermediate period, which could be due to better care, training, feeding, and other management practices in the latter years. However, no consistent trend could be established for the effects of genetic groups and other non-genetic causes on the traits considered.

The objective of the present study was to establish the changes in plasma concentrations of LH, FSH, estradiol 17-beta (E2) and progesterone (P4), as well as to understand their temporal relationships during oestrus in mithun (Bos frontalis). The experiment was conducted on 11 mithuns during third or fourth postpartum oestrous cycle. Since oestrus onset the jugular vein blood samples were collected every 2 h for 72 and 96 h, respectively from the animals without and with standing heat. The LH, FSH, E2 and P4 concentrations were estimated in plasma. The P4 concentration was fluctuated throughout the oestrus period and the average P4 concentration was found significantly (p<0.05) lower on the day of oestrus onset. The multiple rises in LH and FSH concentrations above the basal level in spike like fashion were observed throughout the oestrus period irrespective of the occurrence of standing heat. A significant (p<0.01) gradual increase in the average daily E2 concentration was observed till day 2 following oestrus onset irrespective of the occurrence of standing heat. A significant (p<0.05) simultaneous increase in LH, FSH and E2 concentrations and a transient increase in P4 concentration at approximately the time of standing heat onset were observed. During investigation a definite temporal coupling between LH and FSH rises was absent throughout the oestrus period. The results suggest that (1) the multiple short-duration low-amplitude LH and FSH surges during oestrus may be crucial for the final maturation of ovulatory follicle and subsequent ovulation in mithun; (2) a differential mechanism for controlling LH and FSH secretions probably exists in mithun.