Farmers´ sense of the biological impact of extreme heat and seasonality on Swedish high-yielding dairy cows - A mixed methods approach.

Supporting dairy farmers in becoming resilient towards extreme weather requires a broad understanding of the experiences and perceived risks associated with these events from those who undergo them. We used a mixed methods approach to explore national trends of biological consequences on dairy cow udder health and fertility, combined with in-depth farmer conversations around extreme weather events, focusing on heat. The aim is to provide a comprehensive picture of how dairy farmer perceptions, priorities and decision-making are related to the season and extreme weather to identify preventive pathways that can reduce biological costs of heat stress on Swedish dairy cattle during summer. Data collected monthly at cow and farm level between 2016-2019 as part of the Swedish milk and disease recording system confirm seasonal trends and show increased somatic cell counts (SCC) and negatively impacted fertility during summers. In addition, transcriptions of 18 interviews with dairy farmers across the country and seasonal variations of SCC and fertility were thematically analysed. The results suggest that farmers have a broad definition of extreme weather and are aware of the negative impacts. Yet handling of extreme weather events can mainly be classified as reactive. Nevertheless, there are long-term effects on the farm economy, health and herd dynamics. Swedish dairy farmers are currently showing resilience, albeit a fragile one. The capability to ensure sufficient feed production in extreme weather is critical for farm self-perceived resilience. However, acknowledging the long-term biological costs related to fertility, currently not perceived by farmers, has the potential to support proactive planning and improve farm resilience and profitability.

Herd health and reproductive management associated with lamb weight gain and mortality in sub-Saharan drylands-a case from Ethiopia.

Sheep are important for food and livelihood security in sub-Saharan Africa, and maximizing lamb weight gain while minimizing mortality is essential to improve production. Using the Menz sheep breeding villages of Amhara region in Ethiopia as a case study, the weight gain and mortality rate of 208 lambs were monitored during their first 5 months of life. The study was conducted in intervention and control villages, where the intervention villages were part of community-based breeding programmes and had participated in various projects aiming to improve sheep production and management. Multivariable linear regression analysis was conducted to detect associations between weight gain from birth to 1 month, and birth to 5 months, and different lamb and ewe characteristics, farmer education, application of management routines, and presence of village level sheep management interventions. In general, lambs from intervention villages, without certain signs of diseases, whose mothers were 2 years or older, had a body condition score of more than 2 on a 5-point scale, and who originated from flocks where disease prevention strategies had been implemented, had gained more weight. Overall lamb mortality was 6.8% with most deaths occurring before 1 month of age. This study highlights that health interventions in ewes improve lamb survival and weight gain and that the care of lambs during the first month of life is crucial for overall herd productivity.

A systematic literature review of milk consumption and associated bacterial zoonoses in East Africa.

Consumption of unsafe animal-source foods is the major cause of foodborne disease outbreaks in low-income countries. Despite current knowledge of the threat posed by raw milk consumption to human health, people in many countries in East Africa still consume unboiled milk. This literature review explored the association between milk consumption and the occurrence of five milk-borne bacterial zoonoses: brucellosis, salmonellosis, campylobacteriosis, Escherichia coli infections, and tuberculosis. A search for literature published up to 1 October 2021 was conducted through the Web of Science, PubMed, and Scopus databases, using Preferred Reporting Items for Systematic reviews and Meta-Analyses guidelines. The selection process yielded 65 articles describing studies conducted in East Africa 2010-2021, which were carefully scrutinized. The most investigated pathogen was Brucella spp. (54.5%), followed by E. coli (18.2%), Salmonella spp. (12.1%), Mycobacterium spp. (6.1%), and E. coli O157: H7 (6.1%). The most common predisposing factors for potential milk-borne disease outbreaks were consumption of contaminated raw milk, inadequate cold storage along the milk value chain, poor milk handling practices, and lack of awareness of the health risks of consuming unpasteurized milk. Thus, a tailor-made training program is needed for all milk value chain actors to enhance the safety of milk sold in informal markets, and a One Health approach should be applied. Future studies should employ more advanced diagnostic techniques and countries in East Africa should invest in modern diagnostic tools and equipment, both in hospitals and in local rural settings where most cases occur.

Milk fatty acids as indicators of delayed commencement of luteal activity in dairy cows in early lactation.

Excessive mobilization of adipose reserves due to severe negative energy balance in early lactation may be detrimental to dairy cow fertility at individual and herd level. Reproductive efficiency is one of the main factors influencing herd profitability and a strategy for early detection and management of cows with delayed resumption of cyclicity will result higher conception rate, decreased proportion of cows with extended lactation, fewer inseminations per conception and lower culling rates due to reproductive disorders. Using two groups of dairy cows (Holstein n = 37, Swedish Red breed [SRB] n = 49), we investigated potential differences between cows with different commencement of luteal activity (CLA) and the feasibility of using milk fatty acids (MFAs) as predictors of delayed CLA. Milk samples for progesterone analysis were collected twice weekly during the first six weeks in milk. The concentrations of the MFAs C14:0, C16:0, C18:0 and C18:1 cis-9 in milk (g/100 g milk) and in milk fat (g/100 g fat) were analysed by Fourier transform infrared spectroscopy and individual MFA profiles were calculated by weeks in milk. Commencement of luteal activity was defined as the first day with milk progesterone concentrations >3 ng/ml at two successive measurements. The study population was categorized as early (n = 42) or late (n = 44) CLA, using the median value of 21 DIM as the cut-off. Analysis of the data revealed that CLA was correlated with the proportion of some specific MFAs, where cows with delayed CLA had lower IGF-1 (92.9 ± 7.9 vs. 114.1 ± 7.9 ng/ml; p = .05) and C14:0 levels (10.4 ± 0.2 vs. 11.5 ± 0.2 g/100 fat; p < .01) and higher C18:0 (9.6 ± 0.2 vs. 8.9 ± 0.6 g/100 fat; p < .01) and C18:1 cis-9 levels (24.9 ± 0.4 vs. 23.5 ± 0.4 g/100 fat; p < .05). Delayed CLA (mean 34 days) was predictable for approximately 80% of cows based on C18:0 or C18:1 cis-9 concentrations in week 2 postpartum. Overall, MFAs (C18:0 and C18:1 cis-9) as biomarkers were better indicators than beta-hydroxybutyrate or non-esterified fatty acids in early detection of cows with delayed or normal CLA. The MFA concentrations in milk samples from cows in early lactation can thus be used as a non-invasive method to identify cows at risk of delayed CLA, acting as potential biomarkers for future reproductive performance.

Genetic Characterization of Staphylococcus aureus From Subclinical Mastitis Cases in Dairy Cows in Rwanda.

Whole-genome sequencing was carried out on 30 Staphylococcus (S.) aureus isolates from dairy cows with subclinical mastitis from all five provinces of Rwanda. Twenty-five of the isolates produced enough sequence to be analyzed using core genome multilocus sequence typing (cg-MLST). The isolates group into three main clusters. The largest cluster contain isolates of sequence type (ST) 152 (n = 6) and the closely related ST1633 (n = 2). These sequence types have previously mainly been encountered in humans. The isolates of the second-largest cluster belong to ST5477 (n = 5),so far exclusively isolated from cows in Rwanda. The third cluster consists of isolates of ST97 (n = 4), which is a well-known bovine-adapted sequence type. These three clusters were all widespread over the country. Isolates of the usually human-adapted sequence types 1 (n = 2) and 5 (n= 1) were found and a single isolate of ST2430, previously found among humans in Africa. Finally, four isolates of novel sequence types were found: ST7108 (n = 2), ST7109 (n = 1), and ST7110 (n = 1). The blaZ penicillin resistance gene was found in 84% of the isolates and was in all cases corroborated by phenotypic resistance determination. Five (20%) of the isolates carried a tetracycline resistance gene, tet(K) or tetM, and three of these five also displayed phenotypic resistance while two isolates carried a tetM-gene but were yet tetracycline susceptible. Seven (28%) isolates carried the dfrG gene conferring resistance to trimethoprim. Four of these isolates indeed were resistant to trimethoprim while three isolates were sensitive. The str gene conferring resistance to aminoglycosides was found in three isolates; however, none of these displayed resistance to gentamycin. Our data revealed a high diversity of the sequence types of S. aureus isolates from cows with subclinical mastitis in Rwanda. Two major clusters of ST97 and ST5477 are likely to be bovine adapted and cause mastitis while the third cluster of ST152 usually have been found in humans and may signify a recent transmission of these types from human to cows, for example from hand milking. The high prevalence of this sequence type among dairy cows may pose zoonotic threat. The sequence types were widely distributed without any geographic correlation. Penicillin resistance, the most common type of resistance with a prevalence over 80%, but also tetracycline and trimethoprim resistance were displayed by several isolates.

Extenders for alpaca epididymal spermatozoa: Comparison of INRA96 and andromed.

Artificial insemination would be a useful technique for alpaca breeders to use as an aid to breeding to increase fleece quality. The technique, however, is not well developed in alpacas, partly because of the viscous nature of their seminal plasma. Castration conducted for husbandry purposes can provide a source of epididymal spermatozoa to test semen extenders or handling regimens, thus circumventing the problem of the viscous ejaculate. In this experiment, two semen extenders (Andromed and INRA96) developed for other species (bovine and equine, respectively) were tested with alpaca spermatozoa derived from the cauda epididymis. Sperm total motility (mean ± SEM A: 29.1 ± 4.8 % compared with I: 35.4 ± 4.8 %; NS), membrane integrity (A: 58 ± 9% compared with I: 56 ± 9%; NS) and acrosome integrity (A: 65 ± 7% compared with I: 54 ± 7%; NS) were not different between the two extenders. Progressive motility with use of INRA96 was greater after incubating for 30 min than after incubating for 10 min (35 ± 4% vs. 12 ± 4%, respectively; P = 0.03). In conclusion, viable epididymal spermatozoa could be extracted from the castrated organs after overnight transport. There were no differences in sperm quality between the two extenders; therefore, it appears that either extender could be used for alpaca spermatozoa. These results could help in the development of a technique for artificial insemination in alpacas.

MILK Symposium review: Microbiological quality and safety of milk from farm to milk collection centers in Rwanda.

The aim of this study was to generate knowledge on the most important milk quality and safety attributes, including somatic cell count (SCC), total bacterial count (TBC), Escherichia coli, Salmonella, and Brucella spp. antibodies and antibiotic residues in milk in the chain from farm to milk collection center (MCC) in Rwanda. In addition, we investigated farm and management factors associated with high TBC, SCC, and Salmonella counts. Raw milk was sampled at the farm and MCC levels. Milk samples were taken from dairy farms linked to 2 selected MCC in each of the 4 provinces in Rwanda. In total, 406 bulk milk samples from 406 farms and 32 bulk milk samples from 8 MCC were collected and analyzed. Farm milk average SCC varied between 180 × 103 and 920 × 103 cells/mL, whereas average SCC in milk samples at MCC varied between 170 × 103 and 1,700 × 103 cells/mL. The mean milk TBC of different farms per MCC varied between 1.1 × 106 and 1.6 × 107 cfu/mL, whereas in milk samples from different MCC, the mean TBC ranged between 5.3 × 105 and 2.4 × 108 cfu/mL. The high TBC in milk from MCC suggests proliferation or recontamination of milk by bacteria during transportation. Escherichia coli was detected in 35 of 385 farm milk samples and ranged between 5 cfu/mL and 1.1 × 104 cfu/mL, whereas in milk samples from the MCC, it was detected in 20 out 32 samples varying between 5 cfu/mL and 2.9 × 103 cfu/mL. Overall farm prevalence of Salmonella in milk samples was 14%, but no milk samples from MCC were positive for Salmonella. Five out of 22 bulk milk samples from different MCC were positive for Brucella spp. antibodies, but no Brucella antibodies were detected in milk samples from farms. The prevalence of antibiotic residues as detected by the Delvotest SP NT (DSM, Delft, the Netherlands) was low: 1.3% in farm milk samples and undetected in MCC milk samples. Lack of a separate milking area was associated with high TBC, whereas offering of supplemental feeds, keeping data of past diseases, and an unhygienic milking area were associated with high SCC. Lack of teat washing before milking was the only factor associated with Salmonella contamination of milk at the farm level. This study indicated high TBC and SCC of milk samples at the farm and MCC levels, which indicates both microbial contamination of milk and poor udder health in dairy cows. Presence of E. coli, Salmonella, and Brucella antibodies in milk was common, but finding antibiotic residues in milk was uncommon.

A cross sectional study of prevalence and risk factors associated with subclinical mastitis and intramammary infections, in dairy herds linked to milk collection centers in Rwanda.

The objective of this cross-sectional study was to investigate prevalence, causative udder pathogens and their antimicrobial resistance (AMR), as well as cow and herd risk factors associated with subclinical mastitis (SCM = cows with at least one udder quarter with california mastitis test (CMT) score > 2) and intramammary infections (IMI) caused by Staphylococcus(S.) aureus or Non aureus staphylococci (NAS) in dairy cows linked to Milk Collection Centers (MCCs) in Rwanda. Screening for SCM with the CMT was done on 572 cows from 404 herds linked to two MCCs in each of four provinces. Milk from udder quarters with a CMT score ≥3 (scale 1-5) was sampled for bacteriological analysis. Antimicrobial resistance was evaluated in 60 selected S. aureus isolates. Multivariable mixed effect and ordinary logistic regression analyses were performed to identify cow and herd level risk factors associated with SCM, NAS or S. aureus IMI in cows. The prevalence of SCM was 37.3 % at quarter level and 62.0 % at cow level. Bacteria were isolated 73.7 % of the cultured milk samples, whereas 23.3 % were culture-negative and 3.0 % were contaminated. Staphylococcus aureus and NAS were the most prevalent pathogens, representing more than half of all bacteriological findings. Staphylococcus chromogenes and S. epidermidis were the most prevalent NAS species identified. Of the S. aureus strains 83.3 % were resistant for penicillin, 100 % for clindamycin and 20 % tetracycline. The risk factor analysis showed that an increased stage of lactation, dirty udder and legs in single cow herds and lack of calf suckling the dam, dirty udder and legs and lack of feeding cow after milking in multiple cow herds were significantly (P < 0.05) associated with an increased odds of SCM. Similarly, increasing stage of lactation in single cow herds, and housing cows in individual cattle kraal or on earthen floor and hand washing between cows during milking in multiple cow herds were associated with increased odds for NAS IMI. Poor hygiene of milking area in single cow herds and absence of foremilk stripping in multiple cow herds were significantly (P < 0.05) associated with increased odds for S. aureus IMI in cows. In conclusion, SCM prevalence was high across MCCs. The majority of identified pathogens were contagious in nature and they exhibited resistance to penicillin. Control of the identified risks factors and improved biosecurity through adoption of best practices, and farmer training could contribute to lowering SCM prevalence in Rwanda.

An observational study of the dry period length and its relation to milk yield, health, and fertility in two dairy cow breeds.

This study evaluated associations between dry period length (DPL) in dairy cows and energy-corrected milk (ECM) production, health, and fertility, using a dataset involving 78 577 lactations in cows of two breeds, Swedish Red (SR) and Swedish Holstein (SH). Herds with at least 20 cows and at least 20 % purebred SR and at least 20 % SH during the year 2015/2016 were included in the dataset. Cows were divided into parity groups 2 and 3 or older and DPL was classified into six 10-day groups (30-39 to 80-89 d). Associations between DPL and the outcome variables were assessed with linear and generalized linear mixed models. Production of ECM at the first three test milkings and the 305-d lactation was highest in the 50-59 d and 60-69 d DPL groups, and lowest in the 30-39 d and 80-89 d groups (p < 0.001). Mastitis and somatic cell count (SCC) did not differ between DPL groups, except at the third test milking, where cows in the 40-49, 50-59, and 60-69 DPL groups had lower SCC (p < 0.001) than cows in the 70-79 and 80-89 d groups. The odds ratio (OR) for retained placenta was 1.9 for cows with 30-39 d DPL relative to cows with 60-69 d DPL. Cows in DPL groups 40-49 d and 50-59 ha d the lowest odds of culling (OR=0.89 and 0.92). The OR for culling in cows with DPL 70-79 d and 80-89 was 1.2 and 1.4, respectively, relative to cows with 60-69 d DPL. The conception rate at first insemination did not among between DPL groups. The SR cows produced less ECM than the SH cows, but had lower SCC and lower OR for mastitis, retained placenta, puerperal paresis, and culling within 90 DIM. Although DPL <40 days or>80 days reduced milk production in early lactation and the 305-d lactation, the estimated additional milk production in the preceding lactation in cows with short DPL compensated for this reduction. A DPL of 40 days did not affect health and fertility in comparison with a conventional 60 d DPL, while a DPL of >70 d was unfavorable due to low milk production, increased risk of culling, and decreased fertility. The response to DPL was not affected by breed or parity.

Diseases and causes of death among alpacas in Sweden: a retrospective study.

BACKGROUND: Due to increasing popularity in Sweden during the last decade, alpacas are frequently encountered by practising veterinarians and pathologists. Knowledge regarding their health and diseases under Swedish conditions is, however, limited. OBJECTIVES: To improve knowledge about the health of alpacas in Sweden by collecting information on diseases and health status. DESIGN: A retrospective study was made of 93 necropsies conducted on alpacas in Sweden during the period 2001-2013. SETTING: Data were obtained from the two major veterinary pathology centres in Sweden. The alpacas were hobby or farm animals and they were submitted by veterinarians in local practices or at a national animal healthcare organisation. RESULTS: The digestive system was most frequently affected (29 per cent), with parasitic gastroenteritis (17 per cent) and hepatic disease being especially prevalent (15 per cent fascioliasis and 7 per cent hepatitis). Cardiovascular conditions (9 per cent), systemic diseases (7 per cent) and perinatal deaths were also common, including abortions (10 per cent) and fatal septicaemia (4 per cent). Wasting/emaciation was a frequent finding (26 per cent). Other diagnoses included dermatitis (8 per cent), diseases of the central nervous system (8 per cent), traumatic injuries (7 per cent), neoplasia (5 per cent), pneumonia (5 per cent) and nephritis (3 per cent). CONCLUSIONS: This study identified areas of concern regarding diagnostic and pathological procedures, for which specific measures have been recommended. One particular cause for concern was the number of deaths from emaciation in weanling alpacas during late winter or early spring. For adult alpacas, infectious and non-infectious causes of death were approximately equally frequent. Many of the diseases were considered clinically acute but pathology often showed them to be chronic conditions that had eventually deteriorated and presented as acute cases in the late stages. This study revealed similarities in the health/disease status reported in other European countries and in North America. The results can be used by alpaca keepers and veterinary practitioners to improve management, diagnosis and treatment of alpacas.

Aetiology and prevalence of subclinical mastitis in dairy herds in peri-urban areas of Kigali in Rwanda.

The aim of this cross-sectional study was to evaluate the prevalence of subclinical mastitis (SCM) and associated risk factors in dairy cows in peri-urban areas of Kigali, Rwanda, and identify causative udder pathogens. A sample of 256 cows from 25 herds was screened with the California Mastitis Test (CMT), and udder quarters with CMT score ≥ 3 (scale 1-5) were milk sampled for culture and final bacteriological identification with matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS). All resultant staphylococci species were tested for beta-lactamase production with the clover leaf method. In parallel, herd bulk milk somatic cell count (SCC) of each herd was analysed using a portable device, the DeLaval cell counter. The prevalence of SCM was 43.1% at quarter level and 76.2% at cow level based on CMT test. Multiparous, Holstein cows were 2.50 (C.I = 1.32-4.71) and 10.08 (C.I = 1.54-66.13) times more likely to contract SCM infection than primiparous animals or cows of other breeds, respectively. The median and mean SCC of all herds were 1108 × 103 cells/mL and 1179 × 103 cells/mL, respectively. The most prevalent pathogens were non-aureus staphylococci (NAS; 40.2%) followed by Staphylococcus aureus (22%) and less prevalent pathogens (6%). Samples with no growth or contamination constituted 30.4% and 1.4% of the diagnoses, respectively. The most prevalent species within NAS were S. epidermidis (38.2%) followed by S. sciuri (19.5%), S. chromogenes (9.8%), and nine less prevalent NAS species (32.5%). Out of 209 staphylococci isolates, 77% exhibited beta-lactamase production. The study shows that there is high prevalence of SCM and high herd bulk milk SCC in herds in Kigali, indicating udder health problems in dairy cows. Additionally, beta-lactamase production among staphylococci species was common. Improved milking hygiene and application of biosecurity measures, or a complete mastitis control plan, is required to lower the prevalence of SCM and minimize the spread of pathogens among dairy cows.

DNA methylation pattern of bovine blastocysts associated with hyperinsulinemia in vitro.

Insulin functions as a regulator of metabolism and plays an important role in reproduction. Hyperinsulinemia is often observed in patients with obesity and diabetes type 2 and is known to impair fertility, but the underlying molecular mechanisms are only partly understood. Metabolic programming through epigenetic mechanisms such as DNA methylation during embryonic development can lead to health implications for the offspring later in life. Our aim was to study the potential effect of hyperinsulinemia on gene expression and DNA methylation of embryos by adding insulin (0.1 µg/ml = INS0.1 or 10 µg/ml = INS10) during in vitro oocyte maturation by using the EmbryoGENE DNA methylation array for a study of the bovine epigenome. Our results showed significant differences between blastocysts originating from insulin-treated oocytes compared with untreated control blastocysts. In total, 13,658 and 12,418 probes were differentially methylated (DM) in INS0.1 and INS10, respectively, with an overlap of 3,233 probes in the DM regions (DMR) for both insulin groups. Genes related to pathways such as lipid metabolism, growth and proliferation, mitochondrial function, and oxidative stress responses were influenced at both the epigenetic and transcriptomic levels. In addition, imprinted genes and genes with functions in the epigenetic machinery were among the DMRs. This study identified DMRs correlated to differential expression of genes involved in metabolic regulation and should help to improve our knowledge of the underlying molecular mechanisms of metabolic imbalance.

Lipid profile of bovine blastocysts exposed to insulin during in vitro oocyte maturation.

Insulin is a key hormone with important functions in energy metabolism and is involved in the regulation of reproduction. Hyperinsulinaemia is known to impair fertility (for example, in obese mothers); therefore, we aimed to investigate the impact of elevated insulin concentrations during the sensitive period of oocyte maturation on gene expression and lipid profiles of the bovine Day-8 embryo. Two different insulin concentrations were used during in vitro oocyte maturation (INS10=10µgmL-1 and INS0.1=0.1µgmL-1) in order to observe possible dose-dependent effects or thresholds for hyperinsulinaemia in vitro. By investigating gene expression patterns by an mRNA microarray in combination with lipid profile analysis by desorption electrospray ionisation-mass spectrometry (DESI-MS) of embryos derived from insulin-treated oocytes, we gained further insights regarding molecular responses of embryos to insulin provocation during the first days of development. Lipid metabolism appeared to be influenced on multiple levels according to gene expression results but the profiles collected in positive-ion mode by DESI-MS (showing mostly ubiquinone, cholesteryl esters and triacylglycerols) did not differ significantly from controls. There are parallels in follicular development of ruminants and humans that make this bovine model relevant for comparative research on early human embryonic development during hyperinsulinaemia.

Insulin during in vitro oocyte maturation has an impact on development, mitochondria, and cytoskeleton in bovine day 8 blastocysts.

Insulin is a key metabolic hormone that controls energy homeostasis in the body, including playing a specific role in regulating reproductive functions. Conditions associated with hyperinsulinemia can lower developmental rates in bovine in vitro embryo production and are linked to decreased fertility in humans, as in cases of obesity or type 2 diabetes. Embryo quality is important for fertility outcome and it can be assessed by choosing scoring standards for various characteristics, such as developmental stage, quality grade, cell number, mitochondrial pattern or actin cytoskeleton structure. Changes in the embryo's gene expression can reflect environmental impacts during maturation and may explain morphological differences. Together with morphological evaluation, this could enable better assessment and possibly prediction of the developmental potential of the embryo. The aim of this study was to use a bovine model to identify potential gene signatures of insulin-induced changes in the embryo by combining gene expression data and confocal microscopy evaluation. Bovine embryos were derived from oocytes matured in two different insulin concentrations (10 µg mL-1 and 0.1 µg mL-1), then stained to distinguish f-Actin, DNA and active mitochondria. The total cell number of the embryo, quality of the actin cytoskeleton and mitochondrial distribution were assessed and compared to an insulin-free control group. A microarray-based transcriptome analysis was used to investigate key genes involved in cell structure, mitochondrial function and cell division. Our results indicate that insulin supplementation during oocyte maturation leads to lower blastocyst rates and a different phenotype, characterised by an increased cell number and different actin and mitochondrial distribution patterns. These changes were reflected by an up-regulation of genes involved in cell division (MAP2K2; DHCR7), cell structure (LMNA; VIM; TUBB2B; TUBB3; TUBB4B) and mitochondrial activation (ATP5D; CYP11A1; NDUFB7; NDUFB10; NDUFS8). Taken together, we hypothesise that the increased proliferation in the insulin-treated groups might impair the developmental potential of the embryos by inducing metabolic stress on the molecular level, which could be detrimental for the survival of the embryo.

Bovine herpes virus type 4 alters TNF-α and IL-8 profiles and impairs the survival of bovine endometrial epithelial cells.

Bovine herpes virus type 4 (BoHV-4) can be transmitted by contaminated semen to cows at the time of breeding and may cause uterine disease. The aim of this study was to characterize the susceptibility of bovine endometrial epithelial cells (bEEC) to BoHV-4 by using an in vitro model. When bEEC were challenged with different multiplicity of infection (MOI; from 0.001 to 10) of BoHV-4 for 6days, a significant decrease in cell survival with increasing MOI was observed. The bEEC were subsequently challenged with BoHV-4 MOI 0.1 for 7days. During the first 4days, numbers increased in a similar way in controls and infected group (p<0.01 when compared to Day 0). After Day 4, numbers of live cells in infected samples decreased when compared to controls and were lower than control at Day 7 (p<0.01). From titration and qPCR, increasing number of viral particles was observed from Day 1, and reached a plateau at Day 5. Concentrations of IL-8 increased with time and were higher in supernatants from infected cells than in controls (p<0.0001). TNF-α concentrations presented similar profile as cell survival ones. In conclusion, the survival of bEEC was strongly impaired by BoHV-4 infection in a time and dose dependent manner and supernatant cytokine profiles were altered. This information supports BoHV-4 implication in clinical cases of uterine diseases and the existence of a risk of BoHV-4 transmission from infected males through animal breeding.

A longitudinal cohort study of acute puerperal metritis cases in Swedish dairy cows.

BACKGROUND: Acute puerperal metritis affects cows during the early postpartum period and causes fever, fetid vaginal discharge and general depression. The disease is severe and treatment with antimicrobials is often required. This study followed 79 Swedish dairy cows with acute puerperal metritis with registered treatment and outcome in terms of recovery. Bacteria isolated from the uterus and their susceptibility to penicillin were studied. Clinical cases were assigned by participating practitioners who examined the cows, performed uterine swab sampling, decided treatment and provided information about cow health and calving conditions. Fertility and culling data were collected from the official Swedish milk and health recording scheme. Recovery from disease was defined in four levels; as a cow that survived 1 or 4 months, was inseminated and subsequently became pregnant. Intervals from dates of first and latest calving to insemination date were studied. RESULTS: The most common bacterial findings were a mixed culture of Escherichia coli and bacteria such as Gram positive cocci, Fusobacterium necrophorum, Clostridium spp. or Trueperella pyogenes. The Gram positive cocci, Pasteurella spp. and F. necrophorum were generally susceptible to penicillin. The majority of cows (70%) were treated with penicillin in accordance with the Swedish policy on treatment of metritis while 19% were treated with tetracycline and 8% were not treated with antimicrobials. Recovery rates were similar between treatments. Besides "calving to last insemination" interval (CLI) that was 5 days shorter than the national mean, fertility was slightly reduced compared to national means. "Calving to first insemination" interval (CFI) was 4 days longer than national mean and number of inseminations/cow increased from 1.9 to 2.1. Escherichia coli culture positive cows did not become pregnant to the same extent as cows without E. coli in the uterus (P = 0.046). Twin births resulted in a longer CFI (P = 0.034). The culling rate was generally high (42% within 300 days after occurrence of metritis), though death associated with acute disease was low (6%). CONCLUSIONS: Escherichia coli was the most common bacterial pathogen isolated from cases of acute puerperal metritis in the present study. This bacterium is inherently resistant to penicillin, but although most cows were treated with penicillin, death due to acute disease was low and recovery and final fertility results were acceptable. In times of emerging antimicrobial resistance and demand for prudent antimicrobial use, we suggest that penicillin is a "good enough" choice if antimicrobial treatment of acute puerperal metritis is needed.

Insulin concentrations used in in vitro embryo production systems: a pilot study on insulin stability with an emphasis on concentrations measured in vivo.

BACKGROUND: Insulin has been used as a stimulatory factor for in vitro cell culture since many years. Even for routine in vitro embryo production (IVP), insulin is added to the media during different steps. There is a strong difference in concentrations used in vitro compared to what is measured in vivo in follicular fluid or serum. We performed a pilot study on insulin stability to explain possible reasons for that variation. RESULTS: We measured insulin concentrations before and after bovine oocyte maturation in an experiment by using a quantitative ELISA (Mercodia bovine insulin ELISA immunoassay) and found that concentrations were stable up to 22 h of incubation. We compared our results with eleven in vivo studies measuring insulin in either serum or follicular fluid and nine IVP-protocols using insulin. In all studies, in vitro concentrations were much higher compared with those found physiologically in vivo. Limited knowledge is available concerning the different activity and stability of insulin in vitro versus in vivo. CONCLUSIONS: The concentrations of insulin used in vitro are quite high in comparison to physiological concentrations found in serum or follicular fluid. One explanation may be a different stability or activity of insulin in vitro even if we could measure stable concentrations of insulin in our pilot study. More precise dose-effect studies have to be performed to draw clear conclusions about the consequences of the use of such high doses as they might have negative consequences for the developing embryo. Insulin has direct effects on the regulation of the metabolism and could even influence the epigenetic programming of the metabolism with unknown consequences for the offspring later in life.

The impact of body condition after calving on metabolism and milk progesterone profiles in two breeds of dairy cows.

BACKGROUND: Optimal body condition in early lactation is generally accepted as a prerequisite for good reproductive performance. Examination of milk progesterone profiles offers an objective method for characterization of postpartum ovarian activity in dairy cows. The present study investigated the relationship between body condition after calving, some metabolic parameters in blood plasma, and fertility, as reflected by milk progesterone profiles in the two dairy breeds Swedish Red (SR) and Swedish Holstein (SH). RESULTS: Multiparous dairy cows (n = 73) of SR and SH breeds were selected and divided into three groups based on their body condition score (BCS) after parturition. Selected plasma metabolites were determined, milk progesterone profiles were identified and body condition was scored. Over-conditioned cows and atypical progesterone profiles were more common among SR cows. Insulin sensitivity was lower and IGF 1 higher among SR cows. Insulin was positively related to body condition, but not related to breed. CONCLUSIONS: Atypical progesterone profiles were more common and insulin sensitivity lower in SR than in SH cows, but the SR breed had a higher proportion of over-conditioned SR cows. It is reasonable to assume that breed differences in body condition contributed to these results.

Insulin exposure during in vitro bovine oocyte maturation changes blastocyst gene expression and developmental potential.

Metabolic imbalance impairs fertility, because changes in concentrations of metabolites and hormones in the blood and follicular fluid create an unfavourable environment for early embryonic development. Insulin is a key metabolic hormone known for its effects on fertility: insulin concentrations are increased during energy balance disturbances in diabetes or metabolic syndrome. Still, insulin is frequently used at supraphysiological concentrations for embryo in vitro culture with unknown consequences for the developmental potential of the offspring. In the present study we investigated the effects of insulin exposure during in vitro bovine oocyte maturation on developmental rates, embryo quality and gene expression. Supplementation of the maturation media with insulin at 10 or 0.1 µg mL-1 decreased blastocyst rates compared with an insulin-free control (19.8 ± 1.3% and 20.4 ± 1.3% vs 23.8 ± 1.3%, respectively; P < 0.05) and led to increased cell numbers (nearly 10% more cells on Day 8 compared with control; P < 0.05). Transcriptome analysis revealed significant upregulation of genes involved in lipid metabolism, nuclear factor (erythroid-derived 2)-like 2 (NRF2) stress response and cell differentiation, validated by quantitative polymerase chain reaction. To conclude, the results of the present study demonstrate that insulin exposure during in vitro oocyte maturation has a lasting effect on the embryo until the blastocyst stage, with a potential negative effect in the form of specific gene expression perturbations.

Testicular length as an indicator of the onset of sperm production in alpacas under Swedish conditions.

BACKGROUND: The popularity of alpacas (Vicugna pacos) is increasing in Sweden as well as in other countries; however, knowledge about optimal management practices under Swedish conditions is still limited. The wide age range reported when the onset of puberty can occur, between 1 and 3 years of age, makes management decisions difficult and may be influenced by the conditions under which the alpacas are kept. The aim of this study was to find out when Swedish alpacas can be expected to start producing sperm, by using testicular length and body condition score as a more precise indirect indicator than age. RESULTS: This study suggests that animals with a testicular length ≥3.8 cm would be producing sperm; however, if it is crucial to know that there is no sperm production for management purposes, the threshold level for testicular length used to differentiate between sperm-producing and non-sperm producing animals should be ≤1.6 cm instead. If only one variable is considered, testicular length appears to better than age alone to predict sperm production. Body condition score together with testicular length explains the individual onset of puberty and better guide management recommendations. CONCLUSIONS: Using a combination of these parameters (testicular length, body condition score and age) as a tool for decision making for alpaca husbandry under Swedish conditions is suggested.