First Production of Larvae Using Cryopreserved Sperm: Effects of Preservation Temperature and Cryopreservation on European Eel Sperm Fertilization Capacity

Sperm cryopreservation is a useful tool in captive fish reproduction management, that is to synchronize gamete production, especially in the case of species as the European eel, where the time of female spawning readiness is unpredictable. Several protocols to cryopreserve sperm of this species have been described, but until recently fertilization trials were not feasible. This study evaluated the effect of cold storage of diluted sperm prior to fertilizations and tested whether a previously defined protocol for European eel sperm cryopreservation can be successfully applied in fertilization trials to produce viable offspring. In our experiment, the sperm motility was evaluated after the extraction and the best samples were selected and pooled. Until stripping of eggs and fertilization, diluted sperm samples were maintained at either 4 or 20°C, or cryopreserved, following existing protocols. Fertilization of two egg batches was attempted. Diluted sperm caused a similar percentage of fertilized eggs and a similar number of embryos and larvae, independently of storage temperature (4 or 20°C). The cryopreserved sperm resulted in a lower percentage of fertilized eggs, but embryos developed and a few larvae (‘cryolarvae’) were obtained 55 h after fertilization in one of the two egg batches. This result evidences that the tested cryopreservation protocol is applicable for eel reproduction management, although improvements will be required to enhance fertilization success.     

Changes in Testicular Interstitial Connective Tissue of Hamsters (Mesocricetus auratus) During Ageing and After Exposure to Short Photoperiod

The testicular interstitium of Syrian hamster (Mesocricetus auratus) was studied during ageing and in testicular regression after exposure to a short photoperiod, in relation to the interstitial cells and their connective tissue. This tissue was assessed histochemically using Masson's trichrome technique and the expression of Heat Shock Protein 47 (HSP‐47) and collagen IV (α5) was assessed in Leydig cells. Finally, an ultrastructural analysis of some cells of the testicular interstitium was made. Leydig cells were positive for HSP‐47 and collagen IV (α5). Ageing did not change the parameters studied while the short photoperiod altered the synthetic activity of Leydig cells. The positivity index of these cells for HSP‐47 was significantly higher in the regressed testis, but was lower for collagen IV (α5). During ageing no change were observed. Ultrastructural Leydig cells showed a discontinuous basal lamina that did not change during ageing. The basal lamina was not identified in Leydig cells regressed by exposure to a short photoperiod. In conclusion; the intertubular connective tissue suffers little change with age. By contrast, in the testis regressed after exposure to a short photoperiod the studied parameters related to the intertubular connective tissue were altered. These changes are probably related with the low synthetic activity of regressed Leydig cell.     

Practices and opinions of New Zealand beef cattle farmers towards bovine viral diarrhoea control in relation to real and perceived herd serological status

ABSTRACT

Aims: To investigate the seroprevalence of infection with bovine viral diarrhoea (BVD) virus among 75 beef herds and seroconversion in cattle during early pregnancy, and to determine the practices and opinions of farmers towards BVD control and their association with real and perceived herd serological status.

Methods: Blood samples were collected before mating in 75 beef herds across New Zealand from 15 unvaccinated heifers that had delivered their first calf that season. Serum samples were tested for BVD antibodies using ELISA individually, and after pooling samples for each farm. Animals that were antibody-negative were retested at either pregnancy diagnosis or weaning. Farmers were asked to complete a detailed survey about herd demographics, BVD testing and vaccination practices, and opinions towards national BVD control.

Results: Based on the pooled serum antibody ELISA results, there were 28/75 (37%) negative herds, 15/75 (20%) suspect herds, and 32/75 (43%) positive herds. Of 1,117 animals sampled 729 (65.3%) tested negative for BVD virus antibodies; when retested, 47/589 (8.0%) animals from 13/55 (24%) herds had seroconverted. Among 71 famers providing survey responses 11 (15%) believed their herd was infected with BVD, 24 (34%) were unsure and 36 (51%) did not think their herd was infected. Only 19/71 (18%) farmers had performed any BVD testing within the past 5 years and 50/70 (71%) had not vaccinated any cattle for BVD. Support for national BVD eradication programme was strong in 51/71 (56%) respondents, but the biggest challenge to BVD control was considered to be famer compliance. Compared to farmers who did not think their herd was infected, more farmers who thought BVD was present in their herds had previously tested for BVD, would consider testing all replacement calves, and would support establishing a national BVD database; fewer would consider purchasing BVD tested or vaccinated cattle only.

Conclusions and clinical relevance: Only 15% of the beef farmers in this study believed their herds were infected with BVD virus and few of them had undertaken BVD screening. Nevertheless many were supportive of implementing a national BVD control programme. It is likely that the lack of farmer awareness around BVD and the failure of farmers to recognise the potential impacts in their herds are hindering progress in controlling the disease in New Zealand. There are opportunities for New Zealand veterinarians to be more proactive in helping beef farmers explore BVD management options.     

Paratuberculosis sero-status and milk production,SCC and calving interval in Irish dairy herds

The objective of this study was to investigate the impact of paratuberculosis sero-status on milk yield, fat, protein, somatic cell count and calving interval in Irish dairy herds. Serum from all animals over 12 months of age (n = 2,602) in 34 dairy herds was tested for antibodies to Mycobacterium avium subsp. paratuberculosis using an ELISA. Herds were categorised by sero-status into positive, non-negative and negative, where a positive herd contained two or more positive cows, a non-negative herd contained only one positive cow and a negative herd contained no positive cows. Data at animal, parity and herd-level were analysed by multiple regression using general linear models. Positive herds (mean herd size = 129 cows) and non-negative herds (81 cows) were larger than negative herds (72 cows) (P < 0.01). Negative herds had the highest economic breeding index (EBI), while positive herds had the highest estimated breeding value (EBV) for milk yield. There was no significant effect of paratuberculosis sero-status at animal, parity or herd-level on milk yield, milk fat or protein production, somatic cell count score (SCCS) or calving interval. Negative herds tended to have a lower SCCS than positive and nonnegative herds (P = 0.087). This study only examined the effects of paratuberculosis sero-status but did not examine the clinical effects of Johne's disease at the farm or dairy industry levels.     

Toxicity of dimethoate and fenoxycarb to honey bee brood (Apis mellifera), using a new in vitro standardized feeding method

A new in vitro method was devised to assess the effects of pesticides on honey bee brood. The method allowed the quantification of doses ingested by larvae and the assessment of larval and pupal mortality. Larval mortality in control samples was lower than 10%. Two active substances were tested: dimethoate and fenoxycarb. The LD(50) of dimethoate was 1.9 microg larva(-1) 48 h after oral exposure of larvae at day 4. Additional dose-related effects on pupal mortality were noted. After a chronic intoxication, the NOAEC (No Observed Adverse Effect Concentration) for larval mortality at day 7 was 2.5 mg kg(-1), whereas a NOAEC of 5 mg kg(-1) was found at day 22 for delayed effects on the reduction of adult emergence. Fenoxycarb applied at day 4 showed no effect on larvae, whereas emergence of adults was affected at doses higher than 6 ng larva(-1).     

Plasma Concentrations of Pregnancy‐Associated Glycoproteins Measured Using Anti‐Bovine PAG‐2 Antibodies on Day 120 of Gestation Predict Abortion in Dairy Cows Naturally Infected with Neospora caninum

The present study sought to determine: (i) the effects of Neospora caninum infection and twin pregnancy on plasma pregnancy‐associated glycoprotein‐2 (PAG‐2) concentrations throughout pregnancy and (ii) whether plasma PAG‐2 concentrations could predict abortion in N. caninum‐infected cows. The study was performed on a commercial Holstein‐Friesian dairy herd in northeastern Spain and the final data included those recorded in 53 non‐aborting and 19 aborting animals. Blood samples were collected immediately before pregnancy diagnosis (on Days 40, 90, 120, 150, 180 and 210 post‐insemination) in non‐aborting cows or until the time of abortion detection in aborting cows. General lineal models (GLM) repeated measures anova revealed the different behaviour of PAG‐1 and PAG‐2, and significant effects of Neospora seropositivity, cool season and twin pregnancy on plasma PAG‐2 concentrations throughout gestation (between‐subject effects). In addition, based on the odds ratios, the likelihood of abortion increased in Neospora‐seropositive cows (by a factor of 7.0) compared to seronegative animals and decreased in cows with a high plasma PAG‐2 concentration (>4.5 ng/ml) on Day 120 of pregnancy (by a factor of 0.24), compared to the remaining cows. In conclusion, there is a relationship between plasma PAG‐2 concentrations and the risk of abortion in Neospora‐infected dairy cows. Thus, plasma PAG concentrations measured using anti‐boPAG‐2 antiserum on Day 120 of gestation could serve as an indicator of the abortion risk in N. caninum infected animals; values <4.5 ng/ml indicating a high risk of abortion in chronically infected animals.     

An International Delphi Study of the Causes of Death and the Criteria Used to Assign Cause of Death in Bovine Perinatal Mortality

The objective of the present study was to elicit opinion from two groups of veterinarians [subject matter experts and non‐subject matter experts] about the causes of bovine perinatal mortality and the criteria used to assign such causes. The subject matter experts were selected on the basis of their scientific publications or experience of working in a veterinary diagnostic or research laboratory in the area of bovine perinatal mortality. The non‐subject matter experts were self‐selected as cattle veterinarians without particular expertise in bovine perinatology. A total of 74 veterinarians (46 subject matter experts and 28 non‐subject matter experts) from 23 countries responded. The study was conducted using Delphi methodology over seven rounds. Respondents were asked to agree the causes of bovine perinatal mortality and for each cause to agree the supporting diagnostic criteria. There was a close agreement between groups on 16 causes of death apart from intra‐uterine growth retardation (IUGR) and micronutrient imbalances which were accepted by fewer subject matter experts. There was inter‐group consensus on the criteria to diagnose accidents, congenital defects, dystocia, hyperthermia, infections, premature placental separation, prematurity and prolonged calving. There was inter‐group consensus on the criteria to diagnose anoxia, apart from gingival cyanosis; on haemorrhage, apart from haemorrhagic anaemia; on IUGR, apart from organ weights; and on iodine imbalance, apart from goitre and thyroid iodine content. The results from this study highlighted the current lack of standardization of the criteria used to define the cause of death for bovine perinatal mortality and the need for such standardization.     

64, XX,SRY‐negative,Testicular DSD Syndrome in a Lusitano Horse

Here is reported a disorder of sex development found in the Portuguese Lusitano horse breed. The complex genital phenotype included mammary glands, abdominal testes without epididymis, connected through oviducts to pelvic hypoplastic uterine horns and fused vulvar labia majora from which protruded ventrally a penis‐like structure. This structure was presented in a reversed position, the urethral opening placed dorsally in the glans. However, it was functional both for micturition and erection. The horse exhibited female micturition posture and aggressive male‐like behaviour, including flehmen, mounting, thrusting and flagging of the tail. Plasma testosterone concentrations were below detection limits and the genetic evaluation revealed a 64, XX, SRY‐negative karyotype. Surgery consisted in the removal of abdominal gonads followed by amputation of the penis and repositioning of the urethra. This case of reversion between the chromosomal and gonadal sex, associated with mixed anatomical and behavioural phenotype, illustrates that development of the testes may occur in the absence of the SRY gene and that other genetic and cellular pathways leading to gonad differentiation should be investigated.