Standing Laparoscopic Peritoneal Flap Hernioplasty of the Vaginal Rings does not Modify the Sperm Production and Motility Characteristics in Intact Male Horses

Laparoscopic hernioplasty techniques have been developed in the recent years to avoid the recurrence of inguinal hernias and to spare the testicles for breeding purposes in stallions. However, there have been no previous comprehensive and systematic studies of the reproductive outcomes and prognoses for stallions after inguinal hernioplasty. Therefore, the objective of this study was to assess the possible effects of one of these techniques (standing laparoscopic peritoneal flap hernioplasty) on the sperm production and motility characteristics of six healthy stallions that received this procedure based on 1‐year follow‐ups. There were no significant differences in the measured sperm variables (assessments based on the DSO, MOT, PMOT, VSL, VCL and VAP) during 1‐year follow‐ups.     

The seroprevalence of canine respiratory coronavirus and canine influenza virus in dogs in New Zealand

Abstract

AIM: To determine whether canine respiratory coronavirus (CRCoV) and canine influenza virus (CIV) are present in dogs in New Zealand.

METHODS: Serum samples from 251 dogs of varying age, breed and clinical histories were tested for the presence of antibodies to CRCoV and CIV, using indirect fluorescent antibody (IFA) analysis. The population sampled represented a wide geographic area but principally encompassed the central and lower North Island of New Zealand.

RESULTS: Seventy-three of the 251 samples (29%) were seropositive for CRCoV. Dogs <2 years old were less likely to be seropositive for CRCoV than older dogs. None was seropositive for CIV.

CONCLUSIONS: This study revealed the presence of antibodies to CRCoV in dogs in New Zealand. Young dogs are less likely to be seropositive than older dogs, probably due to increased opportunity for exposure to CRCoV over time. Serum antibodies to CIV were not detected in any of the dogs sampled, suggesting that this virus is unlikely to be present in dogs in New Zealand.

CLINICAL RELEVENCE: Canine respiratory coronavirus is present in New Zealand. Although the role of this virus in canine infectious tracheobronchitis has not been fully elucidated, evidence suggests that it may have a causal role in this disease. Veterinarians should consider CRCoV as a differential diagnosis in cases of respiratory disease in dogs in New Zealand. While CIV appears not to be currently present in New Zealand, veterinarians should consider infection with this virus as a differential diagnosis in dogs presenting with respiratory signs.     

Identification of Apoptotic Bodies in Equine Semen

Apoptosis in the testis is required to ensure an efficient spermatogenesis. However, sometimes, defective germ cells that are marked for elimination during this process escape elimination in the testes, giving rise to ejaculates with increased percentages of abnormal and apoptotic spermatozoa and a high percentage of apoptotic bodies. Apoptosis markers in the ejaculate have been associated with low fertility, either in animals or humans. Therefore, the goal of this study was to investigate whether fresh equine semen contains apoptotic bodies [initially named Merocyanine 540 (M540) bodies] and to study the relationship between the quantity of these bodies and cell concentration, the volume of ejaculate, viability and motility. Moreover, we also studied whether the presence apoptotic bodies in fresh semen was related to the resistance of the stallion spermatozoa to being incubated at 37°C or being frozen and thawed. Fresh equine semen was stained with fluorescent dyes such as M540 and Annexin‐V. Active Caspase 3 was studied in fresh semen through Western blotting and immunofluorescence with a specific antibody. Sperm kinematics was assessed in fresh, incubated and thawed samples using computer‐assisted semen analysis, and viability was evaluated with the LIVE/DEAD Sperm Viability Kit. Overall, our results demonstrate for the first time the presence of apoptotic bodies in equine semen. The quantity of apoptotic bodies was highly variable among stallions and was positively correlated with Caspase 3 activity in fresh samples and negatively correlated with the viability and motility of stallion spermatozoa after the cryopreservation process.     

Effect of Overnight Staining on the Quality of Flow Cytometric Sorted Stallion Sperm: Comparison with Tradtitional Protocols

Flow cytometry is considered the only reliable method for the separation of X and Y chromosome bearing spermatozoa in equines. The MoFlo SX DP sorter is highly efficient, allowing the production of foals of the desired sex. However, to achieve acceptable pregnancy rates the currently used protocol requires working with fresh semen obtained close to, or at, the sorting facility. An alternative protocol was tested during two consecutive breeding seasons. Fresh stallion semen was cooled for 20 h, during which staining with Hoechst 33342 took place. On the following day, this sample was flow sorted and compared with spermatozoa from the same ejaculate that had been sexed on the previous day. All sperm parameters evaluated remained unchanged when fresh sorted and refrigerated sorted semen were compared. Pre‐sorting storage at 5°C did not alter sperm velocities nor kinetics, viability or membrane permeability, production of reactive oxygen species, mitochondrial membrane potential or DNA fragmentation index of the sorted sample. The findings open for the possibility of using semen from stallions housed far from the sorting facilities. Processed and stained sperm could be shipped refrigerated on the previous day, sorted and inseminated on the next day.     

Prevalence and spatial distribution of cattle herds infected with Theileria orientalis in New Zealand between 2012 and 2013

AIM: To describe the prevalence and spatial distribution of cattle herds infected with Ikeda and non-Ikeda types of Theileria orientalis in New Zealand between November 2012 and June 2013.

METHODS: Pooled serum samples collected historically between November 2012 and June 2013 were obtained from cattle herds throughout New Zealand. Each pooled sample consisted of approximately 20 individual cattle samples from that herd, and was provided with details of the spatial location of the herd (n=722). DNA from all samples was tested using two quantitative PCR assays for the detection of T. orientalis (all types) and the Ikeda type. The proportion of herds that were positive for T. orientalis and Ikeda type, or that were positive for T. orientalis but negative for Ikeda type (non-Ikeda positive) was determined for different regions of New Zealand.

RESULTS: The highest prevalence of herds infected with Ikeda type was detected in the Northland (33/35; 94%) and Auckland and the Waikato (63/191; 33%) regions. Only 2/204 (1%) herds were positive for the Ikeda type in the South Island. A high percentage of herds that were positive for non-Ikeda types was detected in the Gisborne and Hawkes Bay (23 (95%CI=13–37)%), Auckland and Waikato (22 (95%CI=16–29)%) and Bay of Plenty (24 (95%CI=10–44)%) regions.

CONCLUSIONS AND CLINICAL RELEVANCE: The high prevalence of Ikeda type detected in cattle herds in the Northland, Auckland and Waikato regions represents a risk to naive cattle being introduced into these regions. There is also the potential for resident cattle herds in the Gisborne and Hawkes Bay, Auckland, Waikato and Bay of Plenty regions to experience increased infection with the Ikeda type.

The overall impact experienced by regions will depend on other factors such as the number of herds present and the predominant type of farming, as well as the interplay between tick ecology, cattle immunity and movement patterns of cattle.     

Lectin Histochemistry as a Tool to Identify Apoptotic Cells in the Seminiferous Epithelium of Syrian Hamster (Mesocricetus auratus) Subjected to Short Photoperiod

Lectins have been widely used to study the pattern of cellular glycoconjugates in numerous species. In the process of cellular apoptosis, it has been observed that changes occur in the membrane sugar sequences of these apoptotic cells. The aim of our work was to identify which lectins, out of an extensive battery of the same (PNA, SBA, HPA, LTA, Con‐A, UEA‐I, WGA, DBA, MAA, GNA, AAA, SNA), show affinity for germinal cells in apoptosis, at what stage of cell death they do so and in which germinal cell types they can be detected. For this, we studied testis sections during testicular regression in Syrian hamster (Mesocricetus auratus) subjected to short photoperiod. Several lectins showed an affinity for the glycoconjugate residues of germ cells in apoptosis: Gal β1,3‐GalNAcα1, α‐d ‐mannose, N‐acetylgalactosamine and l ‐fucose. Furthermore, lectin specificity was observed for some specific germinal cells and in certain stages of apoptosis. It was also observed that one of these lectins (PNA) showed affinity for Sertoli cells undergoing apoptosis. Therefore, we conclude that the use of lectin histochemistry could be a very useful tool for studying apoptosis in the seminiferous epithelium because of the specificity shown towards germinal cells in pathological or experimentally induced epithelial depletion models.     

Risk factors for brucellosis seroprevalence of sheep and goat flocks in Spain

Risk factors for ovine and caprine brucellosis in the Ávila region (center of Spain) were evaluated using data from a cross-sectional study of the most important diseases of small ruminants in this Spanish region between 1996 and 1997. Questionnaire data from 56 herds (35 ovine and 21 caprine) were used. Sixteen (29%) flocks (3 caprine and 13 ovine) were brucellosis-seropositive. Overall, 0.7% of sheep and 0.1% of goats were seropositive. Eleven risk factors were studied at the group level by logistic regression using flock brucellosis-status as outcome, and by linear regression using percentage of brucellosis-seropositivity as outcome. Both final models contained the same variables: contact with sheep and grazing in communal pastures as risk factors, and frequency of disinfecting practices as a protective factor.     

Effect of β‐Carotene Supply During Close‐up Dry Period on the Onset of First Postpartum Luteal Activity in Dairy Cows

The aim of this study was to examine the effect of β‐carotene supply during the close‐up dry period on the onset of first postpartum luteal activity in dairy cows. Twelve cows were supplied with 2000 mg of β‐carotene (20 g Rovimix^®β‐Carotene containing 10%β‐carotene; DSM Nutrition Japan K.K., Tokyo, Japan) by oral administration daily from day 21 before expected calving date to parturition. Fourteen cows (control) did not receive β‐carotene supplementation. Blood samples were obtained on days 21, 14 and 7 before expected calving date and on days 1, 7, 14, 21 postpartum. When the plasma progesterone concentration exceeded 1 ng/ml by day 21 postpartum, luteal activity was assumed to have been initiated. The result showed that serum β‐carotene concentrations in the β‐carotene cows were higher than in the control cows during the experimental period (p < 0.01). The number of cows with the onset of luteal activity by day 21 postpartum was 9/12 in the β‐carotene cows and 4/14 in the control cows (p < 0.05). Retinol, certain metabolic parameters and metabolic hormones concentrations did not differ between β‐carotene and control cows. In addition, serum retinol concentration in β‐carotene cows without luteal activity was lower than in β‐carotene cows with luteal activity (p < 0.05), and serum gamma‐glutamyl transpeptidase concentration in β‐carotene cows with luteal activity (p < 0.05) and control cows without luteal activity (p < 0.01) was higher than in control cows with luteal activity. In conclusion, β‐carotene supply during the close‐up dry period may support the onset of luteal activity during early lactation in dairy cows.     

Soil type as a putative risk factor of ovine and caprine paratuberculosis seropositivity in Spain

Relationships between soil type and ovine and caprine paratuberculosis in the Avila region (central Spain) were evaluated using data from a cross-sectional study of the most-important diseases of small ruminants in this Spanish region between 1996 and 1997. Questionnaire data from 61 herds (38 ovine and 23 caprine) and 1451 serum samples (1041 ovine and 410 caprine) were used. Herd paratuberculosis (herds were scored as positive to paratuberculosis if any of the serum samples was positive in an agar-gel immunodifussion) was the outcome of interest, whereas soil type in the municipality where farms were located was the predictor variable. Other variables related to soil and soil usage, and herd size, replacement, main food production and animal species were also introduced into the multivariable logistic regression. The final model contained only two independent variables: the predictor variable soil type (coded as two dummy variables ST-1 and ST-2) and herd size (dichotomized at the highest quartile). The estimated Odds Ratios were 25.9 (95% CI: 1.6, 411) for ST-1 (entisols as soil type) and 3.5 (95% CI: 0.3, 45) for ST-2 (inceptisols as soil type).     

Dexmedetomidine and midazolam interaction is synergistic with isoflurane and additive with halothane in terms of MAC reduction

Dexmedetomidine and midazolam have synergistic interaction for the sedative/hypnotic and analgesic effects. The purpose of this study was to assess the type of interaction between dexmedetomidine and midazolam for the immobilizing effect in terms of MAC reduction of either halothane (HAL) or isoflurane (ISO). Fifty‐six rats were randomly allocated into one of eight groups (n = 7): SAL + HAL group received saline solution and halothane, SAL + ISO group received saline solution and isoflurane, DEX + HAL group received an intravenous continuous infusion of dexmedetomidine (0.25 μg kg^–1minute^–1) and halothane, DEX + ISO group received an intravenous continuous infusion of dexmedetomidine (0.25 μg kg^–1 minute^–1) and isoflurane, MID + HAL group received an intravenous bolus of midazolam (1 mg kg^–1) and halothane, MID + ISO group received an intravenous bolus of midazolam (1 mg kg^–1) and isoflurane, DEX +MID + HAL group received dexmedetomidine (0.25 μg kg^–1 minute^–1), midazolam (1 mg kg^–1) and halothane and DEX + MID + ISO group received dexmedetomidine (0.25 μg kg^–1 minute^–1), midazolam (1 mg kg^–1) and isoflurane. The tail clamp method was used for MAC determination. Heart rate, invasive arterial blood pressure, respiratory rate and rectal temperature were continuously monitored. Arterial blood gases were analyzed at the end of each experiment. Data were analyzed using a one‐way anova and a Tukey‐Kramer test for multiple comparisons. A p < 0.01 value was considered statistically significant. MAC values were adjusted to the barometric pressure at sea level. Control MAC_bar values expressed as mean ± SD were 1.31 ± 0.11% for HAL and 1.46 ± 0.05% for ISO. Percentages of MAC reduction were 72 ± 17% for HAL and 43 ± 14% for ISO in DEX groups, 26 ± 11% for HAL and 20 ± 9% for ISO in MID groups, and 90 ± 5% for HAL and 78 ± 5% for ISO in DEX + MID groups. The interaction between dexmedetomidine and midazolam in terms of MAC reduction can be described as additive with halothane and synergistic with isoflurane.