Microbial composition of goat buck's ejaculates is modified by the process of preparing and storing refrigerated semen doses.

Ejaculates present their own microbiota, and a link between ejaculates' microbiota and sperm quality and fertility exists. With the development of artificial insemination in animal breeding, ejaculates must be manipulated by diluting them with extenders and storing them at temperatures below body temperature. The effects that these processes have on the original semen microbiota have never been studied. This study explores the effects of the protocol for preparing refrigerated goat buck semen doses and storing on seminal microbiota. Semen from six adult goat bucks of the Murciano-Granadina breed (24 ejaculates) was used, cooled to 4 °C in a skimmed milk-based extender, and stored at this temperature for 24 h. Samples were taken in different steps: in the raw ejaculates (ejaculates), after dilution with the refrigeration extender (diluted), immediately after reaching 4 °C (chilled 0 h) and the samples refrigerated at 4 °C and stored at this temperature for 24 h (chilled 24 h). Sperm quality (motility and integrity of plasma and acrosomal membrane, and mitochondrial functionality) was also evaluated. Bacterial 16S rRNA sequencing was used to study the seminal microbiota. Our results indicated that both refrigeration and storage at 4 °C negatively affected sperm quality parameters. Preparing semen doses and their subsequent conservation caused a significant change in the bacterial community structure. Raw ejaculates showed a lower Pielou's evenness index than the other samples (diluted, chilled 0 h and chilled 24 h). Ejaculates also had a lower Shannon's diversity index (3.44) than the diluted semen (4.17) and the semen chilled for 24 h (4.43). Regarding beta diversity, significant differences were detected between ejaculates and the other treatments. Differences were also found in unweighted UniFrac distances between the semen chilled for 0 h and that chilled for 24 h. At the genus level, marked effects of preparing doses and their subsequent conservation were also evident: 199 genera that were absent in ejaculates were found in the semen chilled and stored for 24 h; 177 genera that were present in ejaculates disappeared after 24-h refrigeration. In conclusion, the extender and protocol for preparing refrigerated goat buck semen doses considerably modify microbial ejaculate composition.

Microbiota in Goat Buck Ejaculates Differs Between Breeding and Non-breeding Seasons.

Changes in semen microbiota are associated with alterations to sperm quality and fertility. However, the microbiota from most livestock species has not yet been studied. Goats are seasonal breeders, but semen microbiota has never been described in this species, and it is unknown how seasonality affects it. Our study objective is 2-fold: to describe the microbiota in goat buck ejaculates and to determine if it differs between breeding and non-breeding seasons. Semen from six males of the Murciano-Granadina breed was collected during both seasons. Two replicates were performed per male and season on different days. The microbiota was characterized by genomic sequencing technology. Sperm quality was also evaluated. Repetition was not significant for the studied variables. Sperm velocities were higher for the breeding than for the non-breeding season. The ejaculates from both seasons also differed in the proportion of apoptotic spermatozoa. The five dominant phyla were Firmicutes, Proteobacteria, Fusobacteria, Actinobacteria, and Bacteroidetes during the breeding season and Firmicutes, Proteobacteria, Actinobacteria, Bacteroidetes, and Cyanobacteria during the non-breeding season. The dominant genus during both seasons was Ureaplasma. Differences in microbial community structure (the beta diversity) were found. A decrease in the relative abundance of the genus Faecalibacterium and an increase in the genera Sphingomonas and Halomonas were observed in the ejaculates collected during the breeding season. Sphingomonas and Faecalibacterium abundance favorably and unfavorably correlated with sperm quality, respectively. In conclusion, the semen microbiota from goat bucks varies between breeding and non-breeding seasons, and the microbiota remains stable for 7 days within a season. In addition, the genera Sphingomonas and Faecalibacterium could be possible biomarkers of semen quality in goat bucks. These results contribute to an in-depth understanding of the effects of reproductive seasonality on goat buck ejaculates.

Terminal Crossbreeding of Murciano-Granadina Goats to Boer Bucks: Effects on Reproductive Performance of Goats and Growth of Kids in Artificial Rearing.

The aim of this work was to improve the growth characteristics of Murciano-Granadina (MG) kids through terminal crossbreeding of MG goats to Boer bucks. Four experiments were carried out, using a total of 354 MG goats, half of which were mated with MG bucks (n = 12) and the other half with Boer bucks (n = 12). The kids were raised in artificial rearing until slaughter weight (9 kg). The birth weight and average daily gain were recorded in crossed kids (n = 197 and 145, respectively) and purebred kids (n = 257 and 169, respectively). Crossed kids presented significant differences (p < 0.001) compared to MG purebred kids in birth weight (+ 24%), mortality in artificial rearing (-37%), average daily gain (+32%) and milk powder conversion rate (-16%). However, the reproductive performance rates of MG goats mated with Boer bucks were slightly worse (pregnancy rate: 78.5% vs. 86.6%, p < 0.05; kidding rate: 62.0% vs. 75.7%; p < 0.01; prolificacy: 1.9 vs. 2.1 kids/parturition), especially when the matings took place in non-breeding season (experiments conducted at latitude 38-39° N). It is concluded that the terminal crossbreeding of MG goats to Boer bucks (those not used to produce replacement kids) could be an interesting option for ethical goat production.

Effect of the Refrigeration System on In Vitro Quality and In Vivo Fertility of Goat Buck Sperm.

Cooling goat sperm insemination doses to 4 °C causes a delay in their delivery. However, chilling these doses during the transportation period could expedite their delivery and the insemination process. In this study, an economical and simple apparatus for chilling goat semen doses in itinere was developed, and the in vitro quality and in vivo fertility of these doses were compared with those chilled by means of a programmable water bath in the laboratory at a rate of -0.18 °C/min. Of the tested prototypes, the one that provided an optimal combination of the chilling rate (average of -0.09 °C/min) and time required to reach 4 °C (3 h 45 min) was selected for further testing. Immediately after chilling and 24 h later, the doses chilled in the prototype were determined to be of higher quality than the samples chilled in the programmable water bath. Finally, the kidding rate was similar between the doses chilled in the programmable water bath (61.7% ± 7.1%) and in the prototype (56.1% ± 5.9%). In conclusion, successful chilling of goat sperm doses during transport is possible, thereby accelerating the delivery of insemination doses.

Effect of Sperm Concentration and Storage Temperature on Goat Spermatozoa during Liquid Storage.

The use of cooled semen is relatively common in goats. There are a number of advantages of cooled semen doses, including easier handling of artificial insemination (AI) doses, transport, more AI doses per ejaculate, and higher fertility rates in comparison with frozen AI doses. However, cooled semen has a short shelf life. The objective of this study was to examine the effect of temperature and sperm concentration on the in vitro sperm quality during liquid storage for 48 h, including sperm motility and kinetics, response to oxidation, mitochondrial membrane potential (MMP) and DNA fragmentation in goats. Three experiments were performed. In the first, the effects of liquid preservation of semen at different temperatures (5 °C or 17 °C), durations (0, 24 and 48 h) and sperm concentrations (250 × 106 sperm/mL (1:2 dilution rate), 166.7 × 106 sperm/mL (1:3 dilution rate) or 50 × 106 sperm/mL (1:10 dilution rate)) on sperm motility and kinetics were studied. In the second experiment, the effect of temperature, sperm washing and concentration on sperm motility and DNA fragmentation was studied. Finally, the effect of sperm concentration and duration of storage at 5 °C on sperm motility, response to oxidative stress and MMP was examined. We found that refrigerated liquid storage of goat sperm impaired sperm quality, such as motility, MMP and response to oxidation, as storage time increased; however, sperm DNA fragmentation index was not significantly affected. Liquid storage at 5 °C preserved higher total motility than at 17 °C. Moreover, we observed that the reduction of sperm concentration below 500 × 106 sperm/mL did not seem to improve the quality of spermatozoa conserved in milk-based extender in the conditions tested.

Quantitative estimation of the impact of caprine arthritis encephalitis virus infection on milk production by dairy goats.

This retrospective study investigated milk production losses associated with serological evidence (serostatus) of caprine arthritis encephalitis virus (CAEV) infection over one lactation in 4543 Murciano-Granadina goats from 22 herds in Spain. The seroprevalence of infection was 18%, ranging from 0% to 2% in 11 herds, 7% to 60% in 10 herds and was 100% in one herd. Seropositive does had significantly shorter lactations, produced less milk and milk fat, lactose and dry extract and had higher somatic cell counts than their seronegative counterparts, although differences in milk production between seropositive and seronegative animals were noted between herds. Mixed regression models confirmed the association between CAEV seropositivity and reduced milk production. The adjusted, least squares mean (LSM) test-day milk yield was 10% less in seropositive compared to seronegative does and this difference varied according to lactation number. In contrast, differences in the LSM of milk fat, lactose and dry extract percentages between seropositive and seronegative goats were only between 0.1% and 0.2% and did not increase with lactation number. The findings of this study provide strong evidence that CAEV-infection can be a major cause of reduction in milk yield in goats and its control should be considered as part of dairy goat herd health schemes.

The relationship between infectious and non-infectious herd factors with pneumonia at slaughter and productive parameters in fattening pigs.

This paper explores the relationship between infectious and non-infectious herd factors with the occurrence of pneumonia at slaughter and productive parameters in fattening pigs on 39 fattening herds. A questionnaire was used to obtain environmental and management factors (non-infectious factors). Blood samples and lungs were obtained from 35 pigs in each herd at slaughter. Serological testing was performed for antibodies against three respiratory pathogens (infectious factors): porcine reproductive and respiratory syndrome virus (PRRSV), Mycoplasma hyopneumoniae (Mh) and Aujeszky's disease Virus-gE protein (ADV-gE). Lung lesion classifications were catarrhal-purulent bronchopneumonia (CPBP), pleuropneumonia (PLP) and pleuritis. A mean lesion value (MLV) was calculated for each lesion. ANOVA and logistic regression assessed statistical associations among MLV, average daily gain (ADG) and feed conversion ratio (FCR) (dependent variables) with infectious and non-infectious factors (independent variables). Mh vaccination was associated with a significant decrease in CPBP; high Mh seroprevalences was associated with an increased level of CPBP. FCR was negatively related with high seroprevalences for ADV-gE and Mh. No significant associations were seen for ADG.

Carcass condemnation causes of growth retarded pigs at slaughter.

Condemnation causes of growth retarded pigs were studied in a Spanish abattoir. A total of 513 carcasses out of 6017 (8.5%) were rejected during inspection. The main reasons for condemnation were abscesses, cachexia, catarrhal bronchopneumonia, vertebral osteomyelitis, arthritis, pleuritis, peritonitis and pleuropneumonia. Positive relationships were found between tail lesions and arthritis (OR=5.23) or vertebral osteomyelitis (OR=24.81), while no relationships were found between tail lesions and abscesses. Lower risks were observed among carcasses condemned for cachexia, and were as follows: abscesses (OR=0.18), arthritis (OR=0.32), vertebral osteomyelitis (OR=0.06). Arcanobacterium pyogenes, either alone or in combination with other agents, was the main bacterial species isolated from abscesses, osteomyelitis and arthritis (73.5% of lesions). Direct economical losses associated with condemnation were calculated to be 30,000 Euro.

Vitrification of goat, sheep, and cattle skin samples from whole ear extirpated after death and maintained at different storage times and temperatures.

Proper tissue preservation from a wide range of animals of different species is of paramount importance, as these tissue samples could be used to reintroduce lost genes back into the breeding pool by somatic cloning. We aim to study the temporal and thermal post-mortem limits, tested in rabbits and pigs, within which there will be guarantees of obtaining living skin cells in goat, sheep, and cattle. We also intend to study the effect of vitrification on the ability of ear skin cells, stored at different times and temperatures, to attach to the substratum and grow in vitro after warming. Ears were stored either at 4 degrees C for 12, 252, and 348 h post-mortem (hpm), or at room temperature (22-25 degrees C) for 60 and 96 hpm. In all cases, skin samples from these ears were sorted into two groups: one group was in vitro cultured immediately after storage, and the other group was vitrified after storage and further in vitro cultured. In goat and sheep, no differences in attachment (100%: goat; 90-100%: sheep) or subconfluence (75-100%: goat; 70-100%: sheep) rates were observed between experimental groups. However, in days of culture to reach subconfluence, significant differences between non-vitrified and vitrified groups were observed when ears were stored at 4 degrees C for 12 and 252 hpm. In cattle, with respect to attachment rate, vitrified samples from ears stored at 22-25 degrees C for 60 hpm were different from non-vitrified control group (60 vs. 100%, respectively; P < 0.05). Also, days of culture to reach subconfluence were analysed by a non-parametric Cox Survival Analysis. In general, results from ANOVA and Survival Analysis were similar, because the proportion of censored data was quite low (9%), so the bias when using ANOVA is not too high. In spite of all the above, the lowest survival rates (75%: goat; 70%: sheep; and 40%: cattle) were sufficiently high to enable collection of skin samples from the majority of dead animals and their cryopreservation.