Genetic analysis of fetal loss in Holstein cattle.

Pregnancy loss is recognized as one of the major factors contributing to poor reproductive performance in dairy cattle. Here, we performed a comprehensive genetic analysis of fetal loss, defined as a pregnancy loss that occurs after detection of a viable embryo around 42 d of gestation. The objectives of this study were to reveal (1) whether fetal loss is heritable and, hence, whether it will respond to selection, and (2) to what extent current fertility traits, such as daughter pregnancy rate, are associated with fetal loss. Data consisted of 59,308 confirmed pregnancy or fetal loss records distributed across nulliparous heifers and primiparous and multiparous cows. We defined fetal loss as a binary trait (yes vs. no) or as an ordinal trait (pregnancy maintenance, early fetal loss ≤150 d of gestation, and late fetal loss >150 d of gestation), and we assessed both linear and threshold models. Heritability estimates for fetal loss ranged from 1 to 18%, depending upon parity, trait definition, and statistical model used. Heritability estimates were greater for lactating cows than for nonlactating nulliparous heifers. Threshold models were able to capture more additive genetic variance and, thus, yielded higher heritability estimates than linear models. Notably, fetal loss traits were highly genetically correlated with each other but only weakly correlated with current fertility traits included in the national genetic evaluation. Overall, our study provides evidence that fetal loss is heritable enough to make genetic selection for reducing fetal loss and improving pregnancy maintenance feasible. In addition, our results suggest that fetal loss is largely independent from current traits used to select for cow fertility, and thus current breeding efforts have unfortunately little effect on reducing the incidence of fetal loss.

MILK Symposium review: Improving control of mastitis in dairy animals in Nepal.

Dairy animals are an important source of income, food, and nutritional security, and improvements in the productivity of dairy animals substantially improve the wellbeing of smallholder dairy farmers. As in other developing countries, dairy animals are key for rural livelihoods in Nepal but often suffer from mastitis-a production disease causing economic losses to farmers, challenges to the dairy processing industry, and possible health hazards to consumers. Studies show that the prevalence of subclinical mastitis in Africa and Asia typically exceeds 50%, threatening animal wellbeing, farmers, dairy processors, and consumers. We conducted a study in Nepal to develop a technology training package to control mastitis in dairy animals. Following identification of knowledge gaps, a technology package consisting of (1) developing good husbandry practices, implementing mastitis detection and control technologies; and (2) training technicians and farmers was implemented. A strategy was subsequently established to provide feedback to farmers in dairy cooperatives on the subclinical mastitis status of their cows. The package was applied in the mid-western region of Nepal. Six months after implementation, we observed a reduction in subclinical mastitis prevalence: from 55% (baseline) to 28% (endline; n = 432) in dairy cows and from 78% to 18% (n = 216) in buffalo. These positive study outcomes strongly suggest that the mastitis technology training package should be scaled across smallholder farmers within and beyond Nepal to control mastitis in dairy animals.

Barriers in accessing family planning services in Nepal during the COVID-19 pandemic: A qualitative study.

BACKGROUND: The COVID-19 virus is still with us, and in resource-limited countries, like Nepal, resurgence of a new variant is still a threat. In this pandemic, low-income countries struggle to provide essential public health services, including family planning. This study was conducted to explore what sorts of barriers are faced by women needing family planning services in Nepal during the pandemic. METHODS: This qualitative study was conducted in five districts of Nepal. Telephonic in-depth interviews were conducted with 18 women of reproductive age (18-49 years) who were the regular clients of family planning services. Data were coded deductively using the preexisting themes based on a socio-ecological model (e.g., individual, family, community, and health-facility levels). RESULTS: Individual level barriers included low self-confidence, inadequate knowledge on COVID-19, myths and misconception related to COVID-19, limited access to FP services low priority to SRH services, low autonomy in family and limited financial ability. Family level barriers comprised of partner's support, social stigma, increased time at home with husbands or parents, un-acceptance of family planning services as essential health services, financial hardship due to loss of jobs, and communication with in-laws. Movement restrictions and transportation hindering access, unsecured feeling, violation of privacy, and obstacles from security personnel were the community level barriers and unavailability of preferred choice of contraception, increased waiting time, limited outreach services by community health workers, limited physical infrastructures, the behavior of health workers, stock out of commodities, and absence of health workers were health facility level barriers. CONCLUSION: This study highlighted key barriers faced by women in seeking family planning services during the COVID-19 lockdown in Nepal. Policymakers and program managers should consider strategies to ensure continued availability of the full method mix during emergency, particularly since disruptions may go unnoticed and strengthen the provision of services through alternative service delivery channels to ensure sustained uptake of such services in this sort of pandemic.

Modern contraceptive use among young women aged 15-24 years in selected municipalities of Western Nepal: results from a cross-sectional survey in 2019.

OBJECTIVE: To estimate the modern contraceptive prevalence rate (mCPR) and its predictors among young women aged 15-24 years. DESIGN: Cross-sectional analysis of Adolescent Youth Project baseline survey. SETTING: 29 municipalities within Lumbini Province and Sudurpaschim Province in Western Nepal. PARTICIPANTS: 683 young women aged 15-24 years who were living in the catchment area of the selected 30 private OK network health facilities at the study sites from November to December 2019 and who provided informed consent or assent. OUTCOME MEASURE: mCPR among young women aged 15-24 years. RESULTS: The mean age of the respondents was 19 years, 61.7% never had sex and 63.9% were unmarried. The mCPR was 11.9% (95% CI 9.5 to 14.8). Of those who reported using a modern method of contraception, injectables (37.9%) were the most common, followed by male condom (35.9%) and implants (8.8%). Majority (86.4%) of the respondents reported currently not using any method of contraception. In the binary logistic regression analysis, the odds of contraceptive use were higher among women aged 20-24 years (adjusted OR (AOR)=5.50, 95% CI 2.94 to 10.29) and those of Janajati caste/ethnicity (AOR=2.08, 95% CI 1.16 to 3.71), while the odds were lower among women who faced high level of barriers (individual, family/societal, service provider and health facility barriers) to contraceptive use (AOR=0.36, 95% CI 0.14 to 0.98). CONCLUSIONS: The mCPR among young women aged 15-24 years was low but similar to the national level. Sexual and reproductive health programmes aiming to improve the mCPR in this population of young women should consider the reported level of sexual activity. Reaching young women to improve their knowledge and self-efficacy for contraception is critical to ensure they can access contraception when needed. The focus should be on reaching not just young women but also key influencers and service providers and making health facilities adolescent-friendly to reduce barriers to contraceptive uptake and to realise self-efficacy.

Medical abortion drug dispensing practices among private pharmacy workers in Nepal: A mystery client study.

BACKGROUND: Pharmacies are the first point of contact for women seeking medical abortion (MA) and act as important sources of information and referral in Nepal. Over the counter sale of MA drugs is not currently allowed in Nepal. This study aimed to assess the MA drug dispensing practices of pharmacy workers using mystery clients in Nepal. METHODS: A cross-sectional study using the mystery client approach was conducted in 266 pharmacies in September-October 2019. These pharmacies had either received harm reduction training or medical detailing visits. A total of 532 visits were conducted by six male and six female mystery clients. Mystery clients without prescription approached the sample pharmacy and filled out a standard digital survey questionnaire using the SurveyCTO application immediately after each interaction. RESULTS: Pharmacy workers dispensed MA drugs in 35.7% of the visits while they refused to provide MA drugs to the mystery clients in 39.3% of visits. Lack of evidence of prior consultation with a physician (27.5%), referral to other health facilities (21.8%), unavailability of MA drugs in the pharmacy (21.3%) and lack of prescription (16.4%) were the main reasons for refusal. Seventy percent of the pharmacy workers inquired clients about last menstrual period/months of pregnancy while 38.1% asked whether the pregnancy status was confirmed. During 65.1% of the visits, mystery clients were told about when to take the MA drugs while in 66.4% of visits, they were told about the route of drug administration. Similarly, mystery clients were briefed about what to expect during the abortion process in half of the visits, and information about the possible side effects of the MA drug was provided in 55.9% of the visits. Pharmacy workers provided correct information on taking MA drugs to mystery clients in 70.7% of visits. CONCLUSION: Despite legal provision of sale of MA drugs only on prescription, pharmacy workers dispensed MA drug in one out of three visits. As pharmacies are the initial contacts of women for abortion services in Nepal, correct supplementary information through pharmacy workers can be an effective strategy to expand access to quality safe abortion services.

Genetic Evaluations of Stillbirth for Five United States Dairy Breeds: A Data-Resource Feasibility Study.

Genetic selection has been an effective strategy to improve calving traits including stillbirth in dairy cattle. The primary objectives of the present study were to characterize stillbirth data and determine the feasibility of implementing routine genetic evaluations of stillbirth in five non-Holstein dairy breeds, namely Ayrshire, Guernsey, Milking Shorthorn, Brown Swiss, and Jersey. An updated sire-maternal grandsire threshold model was used to estimate genetic parameters and genetic values for stillbirth. Stillbirth data with the birth years of dams from 1995 to 2018 were extracted from the United States national calving ease database maintained by the Council on Dairy Cattle Breeding. The extracted stillbirth records varied drastically among the five dairy breeds. There were approximately 486K stillbirth records for Jersey and more than 80K stillbirth records for Brown Swiss. The direct and maternal heritability estimates of stillbirth were 6.0% (4.5-7.6%) and 4.7% (3.3-6.1%) in Jersey and 6.8% (3.2-10.5%) and 1.1% (0.6-2.9%) in Brown Swiss. The estimated genetic correlations between direct and maternal genetic effects for stillbirth were -0.15 (-0.38 to -0.08) in Jersey and -0.35 (-0.47 to -0.12) in Brown Swiss. The estimated genetic parameters for stillbirth in these two breeds were within close ranges of previous studies. The reliabilities of predicted transmitting abilities in Jersey and Brown Swiss increased substantially, thanks to the substantial increase in available stillbirth data in the past 10 years. The stillbirth records for Ayrshire, Guernsey, and Milking Shorthorn, which ranged approximately between 3K and 12K, are insufficient to implement reliable routine genetic evaluations of stillbirth in these three dairy breeds. Estimated genetic (co)variances and genetic values deviated considerably from the reported ranges of previous studies, and the reliabilities of predicted transmitting abilities were low in these three breeds. In conclusion, routine genetic evaluations of stillbirth are feasible in Brown Swiss and Jersey. However, reliable genetic evaluations of stillbirth in Ayrshire, Guernsey, and Milking Shorthorn require further data collection on stillbirth.

Genes and pathways associated with pregnancy loss in dairy cattle.

Pregnancy loss directly impairs reproductive performance in dairy cattle. Here, we investigated genetic factors associated with pregnancy loss following detection of a viable embryo around 42 days of gestation. The objectives of this study were to perform whole-genome scans and subsequent gene-set analyses for identifying candidate genes, functional gene-sets and gene signaling pathways implicated in pregnancy loss in US Holstein cows. Data consisted of about 58,000 pregnancy/abortion records distributed over nulliparous, primiparous, and multiparous cows. Threshold models were used to assess the binary response of pregnancy loss. Whole-genome scans identified at least seven genomic regions on BTA2, BTA10, BTA14, BTA16, BTA21, BTA24 and BTA29 associated with pregnancy loss in heifers and lactating cows. These regions harbor several candidate genes that are directly implicated in pregnancy maintenance and fetal growth, such as CHST14, IGF1R, IGF2, PSEN2, SLC2A5 and WNT4. Moreover, the enrichment analysis revealed at least seven significantly enriched processes, containing genes associated with pregnancy loss, including calcium signaling, cell-cell attachment, cellular proliferation, fetal development, immunity, membrane permeability, and steroid metabolism. Additionally, the pathway analysis revealed a number of significant gene signaling pathways that regulate placental development and fetal growth, including Wnt, Hedgehog, Notch, MAPK, Hippo, mTOR and TGFß pathways. Overall, our findings contribute to a better understanding of the genetic and biological basis of pregnancy loss in dairy cattle and points out novel strategies for improving pregnancy maintenance via marker-assisted breeding.

Comparison between the Manual Method of Indirect Coombs via Gel Technology and Solid Phase Red Cell Adherence.

Objective: An experiment was conducted to define the principle of the conventional tube technique, gel card technology, and solid-phase technology of blood transfusion. The study also highlights the test reactions and various methods of grading reactions for each technology. It further discusses the automated equipment available for each technology and compares the equipment, tests reactions, procedures, and sensitivity of these techniques. Material and methods: This cross-sectional study was conducted on blood samples of 40 patients with positive indirect antiglobulin test at the Padmashree Diagnostics Center and the Bangalore Medical Services Trust, India. Tube and gel card method and solid-phase red cell adherence assay (SPRCA) were evaluated. Results: The results revealed SRPCA of 1+ in nine samples, 2+ in 15 samples, 3+ in 13 samples and 4+ in three samples, while the manual method yielded 1+ in 14 samples, 2+ in 13 samples, 3+ in 13 samples and 4+ in one sample. Conclusion: Solid-phase red cell adherence assay is more precise and capable of detecting red cell adherence assay than tube method and indirect Coombs test gel technology.

Citric acid/ß-alanine carbon dots as a novel tool for delivery of plasmid DNA into E. coli cells.

Successful delivery of plasmid DNA into the microbial cells is fundamental in recombinant DNA technology. Natural bacterial transformation is limited to only certain species due in part to the repulsive forces between negatively charged DNA and bacterial membranes. Most common method of DNA delivery into bacteria is artificial transformation through heat shock and electroporation. These methods require sophisticated instruments and tedious steps in preparation of competent cells. Transformation by conjugation is also not applicable to all plasmids. Nanoparticles have been used successfully in therapeutics for drug delivery into animal cells. They are starting to gain popularity in plant sciences as novel DNA nano carriers. Despite their promise as tool for DNA delivery, their use in microbial cell transformation has not been reported yet. Here we report the synthesis of carbon dots (CDs) from citric acid and ß-alanine and their use in DNA delivery into E. coli cells. CDs were fabricated using microwave assisted synthesis. Plasmids carrying RFP reporter and ampicillin resistance genes were transferred to bacterial cells and further confirmed using polymerase chain reaction. Our findings indicate that CDs can be used successfully for delivery of foreign DNA of up to 10 kb into E. coli. We have demonstrated the use of ß-alanine/citric acid carbon dots as nanocarriers of DNA into E. coli cells and identified their limitation in terms of the size of plasmid DNA they could carry. Use of these carbon dots is a novel method in foreign DNA delivery into bacterial cells and have a potential for the transformation of resistant organism for which there is still no reliable DNA delivery systems.

Maternal risk factors for underweight among children under-five in a resource limited setting: A community based case control study.

Previous studies conducted in Nepal have not identified the potential maternal risk for underweight among children under-five years of age in resource-poor settings. Therefore, to identify these risk factors for being underweight among children under-five years old, a community-based case-control study was conducted in a rural village in the Chitwan District in Nepal. Cases were defined as children who were diagnosed as underweight based on low weight per age, whereas controls were the children with normal weight for their age. Mothers of 93 cases and 186 controls were invited for an interview to collect the data. More than half of underweight children were female (51.6%) and nearly one third of them (31.2%) were aged 13-24 months. Nearly, 30% of the cases belonged to families in the lowest wealth quintile and 82% of cases were from food insecure families. Logistic regression analysis showed that children of mothers who were illiterate had 1.48 times the odds of being underweight compared to whose mothers were not illiterate (95% Confidence Interval [CI]: 1.53-3.07)). Children whose mother had not completed their postnatal care (PNC) were 3.16 more times likely to be underweight compared to children of mothers who completed PNC (95% CI: 1.24-8.03). The children who received care from other family members besides their mothers were 6.05 times more likely to be underweight (95% CI: 1.44-25.42); the children having mothers who had no income at all had 5.13 times the odds of being underweight (95% CI: 1.27-20.71) and children with diarrhea episodes within one month were 2.09 times more likely to be underweight (95% CI:1.02-4.31) compared to those children without any diarrhea episodes within one month. Women should be encouraged to take care of their children themselves, seek PNC services and take precautions to protect their children from diarrhea. Also, enabling factors such as education and improved income for women can help to reduce malnutrition among children.

Whole Genome Mapping Reveals Novel Genes and Pathways Involved in Milk Production Under Heat Stress in US Holstein Cows.

Heat stress represents a major environmental factor that negatively affects the health and performance of dairy cows, causing huge economic losses to the dairy industry. Identifying and selecting animals that are thermotolerant is an attractive alternative for reducing the negative effects of heat stress on dairy cattle performance. As such, the objectives of the present study were to estimate genetic components of milk yield, fat yield, and protein yield considering heat stress and to perform whole-genome scans and a subsequent gene-set analysis for identifying candidate genes and functional gene-sets implicated in milk production under heat stress conditions. Data consisted of about 254k test-day records from 17,522 Holstein cows. Multi-trait repeatability test day models with random regressions on a function of temperature-humidity index (THI) values were used for genetic analyses. The models included herd-test-day and DIM classes as fixed effects, and general and thermotolerance additive genetic and permanent environmental as random effects. Notably, thermotolerance additive genetic variances for all milk traits increased across parities suggesting that cows become more sensitive to heat stress as they age. In addition, our study revealed negative genetic correlations between general and thermotolerance additive effects, ranging between -0.18 to -0.68 indicating that high producing cows are more susceptible to heat stress. The association analysis identified at least three different genomic regions on BTA5, BTA14, and BTA15 strongly associated with milk production under heat stress conditions. These regions harbor candidate genes, such as HSF1, MAPK8IP1, and CDKN1B that are directly involved in the cellular response to heat stress. Moreover, the gene-set analysis revealed several functional terms related to heat shock proteins, apoptosis, immune response, and oxidative stress, among others. Overall, the genes and pathways identified in this study provide a better understanding of the genetic architecture underlying dairy cow performance under heat stress conditions. Our findings point out novel opportunities for improving thermotolerance in dairy cattle through marker-assisted breeding.

Gene Mapping and Gene-Set Analysis for Milk Fever Incidence in Holstein Dairy Cattle.

Milk fever is an important metabolic disorder that affects dairy cows around parturition. It is associated with a breakdown in the mechanisms of calcium homeostasis, resulting in very low blood calcium levels (hypocalcemia). The main objective of this study was to dissect the genetic basis underlying milk fever incidence in Holstein cattle. Data consisted of 31.6 k producer-recorded lactation incidence records from 15.3 k cows. The analysis included a whole-genome scan and a subsequent gene-set analysis in order to reveal individual genes, genetic mechanisms and biological pathways implicated in the incidence of periparturient hypocalcemia. The association analysis identified at least eight different genomic regions that explain considerable amounts of additive genetic variance for milk fever incidence. Notably, some of these regions harbor genes, such as CYP27A1, CYP2J2, GC, SNAI2, and PIM1, that are directly involved in vitamin D metabolic pathway. Moreover, the gene-set analysis revealed several functional terms, such as calcium ion binding, calcium ion transportation, T cell differentiation, B cell activation, protein phosphorylation, apoptosis, and protein kinase activity, among others, that could be implicated in the development of periparturient hypocalcemia. Overall, this comprehensive study contributes to a better understanding of the genetic control of this complex disease. In addition, these findings may contribute to the development of novel breeding strategies for reducing the incidence of milk fever in dairy cattle.