How Stallion Sperm Age In Vitro? Scenario for Preservation Technologies

Recent findings on the molecular damage occurring in the stallion spermatozoa are reviewed. Mechanisms leading to cell death or survival are briefly overviewed, and recent discoveries on molecular pathways leading to sperm death and sublethal damage are discussed. Increasing the understanding of this particular area may disclose clues to develop new strategies to improve current sperm conservation methods.     

Conjugation and Characterization of Latex Particles with Toxoplasma gondii–specific Immunoglobulin Y Antibodies for Diagnostic Aim and Evaluation Efficiency in In Vitro Culture

Toxoplasma gondii is a parasite that causes severe health problems in the world. Toxoplasmosis, an infection caused by T. gondii, leads to high risk of mortality in patients with immunodeficiency, transplantation, and cancer. Besides that, it causes miscarriages in pregnancy, various abnormalities such as hydrocephalus in infants and congenital diseases. Because the clinical indication of the disease is not specific, it is confused with many diseases, and this leads to the necessity of directly detecting the presence of the toxoplasmosis. Therefore, various diagnostic assays are needed for the diagnosis of the disease. Amongs them, latex agglutination assay is widely used for the detection of specific antibodies or antigens in samples. Latex particles are coated with immunogenic molecules (antigens) to detect antibodies in the blood or used to identify antigens when coated with specific antibodies. In both, aggregation of latex particles results in agglutination. Monoclonal antibodies are often used in latex agglutination assay as in other diagnostic methods. However, monoclonal antibodies can be produced in low quantities at a high cost. Besides, to produce monoclonal antibodies, an experienced staff, a well-equipped cell culture laboratory, a long period of time, and a burdened budget are needed. In recent years, as an alternative to monoclonal antibodies, immunoglobulin Y (IgY) antibodies, which are obtained from chicken eggs, and specifically produced against desired antigenic constructs, have become quite attractive in terms of both low cost and abundant production without requiring infrastructure. In contrast, the latex assay based on IgY antibodies for use in the diagnosis of T. gondii has not been developed. This study aimed to conjugate T. gondii-specific IgY antibodies to latex particles, characterize the particles by Fourier transform infrared spectroscopy, scanning electron microscopy, and spectroscopic methods, and finally demonstrate the interaction with T.gondii parasites in culture with scanning electron microscopy analysis.     

The Phosphodiesterase Inhibitor,Isobutyl-1-Methylxanthine Prevents the Sudden Drop in Cyclic Adenosine Monophosphate Concentration and Modulates Glucose Metabolism of Equine Cumulus–Oocyte Complexes Matured in Vitro

Spontaneous nuclear maturation of mammalian oocytes can occur when physically removed from the ovarian follicle during in vitro oocyte maturation (IVM), largely because of a decrease in cyclic adenosine monophosphate (cAMP) concentration. Modulation of oocyte cAMP during IVM by using phosphodiesterase inhibitors has been shown to maintain elevated oocyte cAMP concentrations and control meiotic resumption of bovine and ovine oocytes. This study determined the effect of inclusion of isobutyl-1-methylxanthine (IBMX) during collection and the first 12 hours of incubation of equine oocytes on cAMP concentration and glucose metabolism of cumulus–oocyte complexes (COCs). Abattoir-derived COCs were collected in aspiration medium with (Asp-IBMX) or without (Asp) IBMX. Cumulus–oocyte complexes were then incubated for 12 hours in IVM medium with (Mat-IBMX) or without (Mat) IBMX, followed by additional 24 hours in Mat medium. The cAMP concentration, glucose consumption, lactate production, and metaphase II rates of the COCs were assessed. Cumulus–oocyte complexes aspirated into Asp-IBMX (62.2 ± 2.6 fmol per COC) medium had higher cAMP concentration than Asp (31.8 ± 2.8 fmol per COC) control group (P < .05). Likewise, at 12 hours of IVM, Mat-IBMX group (33.2 ± 2.1 fmol per COC) had higher cAMP concentration than the Mat group (7.68 ± 0.5 fmol per COC; P < .05). Glucose consumption and lactate production were lower during the first 12 hours of incubation in COCs cultured in Mat-IBMX (P < .05). Isobutyl-1-methylxanthine prevented the rapid drop in cAMP concentration and altered metabolism of glucose by the COC. Preventing the sudden drop in cAMP prevents the premature nuclear maturation of in vitro–matured oocytes causing poor developmental competence.     

Reduced Fertility in High‐yielding Dairy Cows: Are the Oocyte and Embryo in Danger? Part I The Importance of Negative Energy Balance and Altered Corpus Luteum Function to the Reduction of Oocyte and Embryo Quality in High‐yielding Dairy Cows*

Fertility in high yielding dairy cows is declining, and there is increasing evidence to presume that oocyte and embryo quality are major factors in the complex pathogenesis of reproductive failure. In this report we present an overview of possible mechanisms linking negative energy balance (NEB) and deficiencies in oocyte and embryo developmental competence; specifically, in the high producing dairy cow. Changes in follicular growth patterns during a period of NEB can indirectly affect oocyte quality. The endocrine and biochemical changes, which are associated with NEB, are reflected in the microenvironment of the growing and maturing female gamete, and likely result in the ovulation of a developmentally incompetent oocyte. Even after an oocyte is successfully ovulated and fertilized, a full‐term pregnancy is still not guaranteed. Inadequate corpus luteum function, associated with reduced progesterone, and probably also low insulin‐like growth factor concentrations, can cause a suboptimal microenvironment in the uterus that is incapable of sustaining early embryonic life. This may partly account for the low conception rates and the high incidence of early embryonic mortality in high yielding dairy cows.     

Molecular epidemiological and phylogenetic analyses of canine parvovirus in domestic dogs and cats in Beijing, 2010–2013

Fifty-five samples (15.62%) collected from dogs and cats were identified as canine parvovirus (CPV) infection in Beijing during 2010–2013. The nucleotide identities and aa similarities were 98.2–100% and 97.7–100%, respectively, when compared with the reference isolates. Also, several synonymous and non-synonymous mutations were also recorded for the first time. New CPV-2a was dominant, accounting for 90.90% of the samples. Two of the 16 samples collected from cats were identified as new CPV-2a (12.5%), showing nucleotide identities of 100% with those from dogs. Twelve samples (15.78%) collected from completely immunized dogs were found to be new CPV-2a, which means CPV-2 vaccines may not provide sufficient protection for the epidemic strains.     

Is Sperm Freezability Related to the Post‐Thaw Lipid Peroxidation and the Formation of Reactive Oxygen Species in Boars?

The aim of the present study was to determine whether the levels of reactive oxygen species (ROS) substances production and the levels of lipid peroxidation of the sperm membrane were related to the quality that the ejaculates exhibited after cryopreservation in boars. Ejaculates from 42 healthy boars were used in this study and they were cryopreserved with the lactose‐egg yolk extender (LEY). Several sperm quality parameters were assessed by flow cytometry in samples incubated for 30 and 150 min at 37°C after thawing: the percentage of sperm with intact plasma membrane (SIPM), intracellular reactive oxygen substances production through mean of DCF fluorescence intensity of total sperm (mean‐DCF) and the percentage of viable and non‐viable sperm containing oxidized BODIPY (VSOB and NVSOB). In addition, the percentages of total motile (TMS) and progressively motile sperm (PMS) were assessed at the same incubation times with a computer‐assisted sperm analysis system. The classification of the ejaculates into good or bad freezers was performed through hierarchical cluster analysis from SIPM and TMS at 150 min post‐thawing. The ejaculates of those males classified as good freezers exhibited higher (p < 0.05) SPIM, TMS and PMS than the bad freezers, although both groups presented similar (p > 0.05) VSOB, NVSOB and mean‐DCF. Therefore, these results show that lipid peroxidation and the amount of reactive oxygen substances in the sperm after cryopreservation are similar between boars classified as good or bad freezers.     

Distribution of IS629 and stx genotypes among enterohemorrhagic Escherichia coli O157 isolates in Yamaguchi Prefecture,Japan, 2004–2013

Patterns of insertion sequence (IS)629, norV genotype, and Shiga toxin (Stx) genotype distribution were investigated amongst 203 enterohemorrhagic Escherichia coli O157 isolates collected in Yamaguchi Prefecture, Japan, between 2004 and 2013. A total of 114 IS629 patterns were identified; these were divided into eight IS groups (A–H). Ninety isolates carried an intact norV gene, whereas 113 isolates carried a norV with a 204-bp deletion. Other than one isolate from IS group G, all isolates with an intact norV belonged to groups A–F, whereas isolates with a mutant norV belonged to IS groups G and H. Seven stx genotypes were identified, and of those, stx1a/stx2a was predominant (n=105), followed by stx2c (n=32) and stx2a (n=27). The stx1a/stx2a genotype was associated with the mutant norV isolates, whereas isolates with an intact norV had the stx2c genotype. Therefore, certain combinations of IS type and stx genotype appear to be more frequent among O157 clades which may be useful for detection of predominant subtypes in the interest of public health.     

Effect of dietary digestible lysine level on growth performance,blood metabolites and meat quality of broilers 23–38 days of age

This study was done to evaluate the effects of different dietary digestible lysine (dig Lys) levels on growth performance, blood metabolites, carcass and breast yield, and breast meat quality of broilers 23–38 days of age. Three hundred 23-day-old Cobb-500 male broiler chickens were allocated to a completely randomized design with five treatments (finisher diet containing 0.88%, 0.94%, 1.00%, 1.06% and 1.12% dig Lys) and six replicates of 10 birds each. Feed intake (FI) was not affected by different dietary dig Lys levels. Weight gain (WG) linearly increased and feed conversion ratio (FCR) linearly decreased with an increasing dietary dig Lys levels. With increasing dietary dig Lys levels, carcass and breast yield and breast meat hue angle (h^*) linearly increased, but abdominal fat decreased quadratically, whereas breast meat lightness (L^*) and ether extract composition linearly decreased. Dietary dig Lys levels did not show any significant effect on serum metabolites, breast meat cooking loss and water holding capacity. Based on the linear broken-line regression models, the weight gain, feed conversion ratio and breast yield were optimized when dietary dig Lys levels were 0.95%, 1.01% and 1.02% respectively. It is concluded that Lys requirements vary according to what productive parameter is taken for optimization. A minimum of 1.02% dig Lys concentration in the finisher diet is suggested to optimize breast yield, feed efficiency and performance in broiler chickens.     

The Concentrations of LH,FSH, Oestradiol-17β and Progesterone in the Blood Plasma of the Female Bactrian Camel (Camelus bactrianus) before and after Intramuscular Injection of Seminal Plasma

The concentrations of LH, FSH, oestradiol-17beta and progesterone in the blood plasma of the female Bactrian camel (Camelus bactrianus) before and after intramuscular injection of seminal plasma.     

Arrhythmogenic right ventricular cardiomyopathy in Bulldogs: Evaluation of clinical and histopathologic features,progression, and outcome in 71 dogs (2004–2016)

ObjectivesThis study aimed to characterize the clinical and histopathological features of arrhythmogenic right ventricular cardiomyopathy (ARVC) in English Bulldogs, American Bulldogs, and Bulldog-type mixed breed dogs and assess affected Bulldogs for a striatin gene mutation previously reported in Boxers with ARVC.AnimalsSeventy-one Bulldogs fit the inclusion criteria. Genetic analysis was performed on five dogs. Cardiac post-mortem evaluations were performed on two dogs.MethodsMedical records from a single veterinary cardiology group (CVCA) were retrospectively evaluated. Tissue and blood samples were submitted for histopathological analysis and genetic testing in select patients.ResultsPresenting complaints included syncope (38%), arrhythmia (81.7%), or murmur (34.2%) documented on examination. On presentation, congestive heart failure (CHF) was diagnosed in 22 (31%) dogs, and 58 (81.7%) had ventricular arrhythmias. On bivariable analyses, the two-dimensional (2D) left atrial-to-aortic root ratio (LA:Ao) was the only prognostic variable significantly associated with survival time. Dogs with 2D LA:Ao below the mean (1.41) had longer median survival to all-cause mortality (12 months; 95% confidence interval [CI] six–15 months) than those with 2D LA:Ao above the mean (four months; 95% CI two–six months; p=0.0384). Most dogs (54%) died from cardiac disease, with 42.1% experiencing sudden death. The median time from diagnosis to cardiac death was four months.ConclusionsArrhythmogenic right ventricular cardiomyopathy affects Bulldogs with both arrhythmogenic and dilated-type phenotypes. Despite variable arrhythmia severity and predominantly right-sided involvement in many dogs, an increase in left atrial size was the only significant predictor of mortality in this sample of dogs.     

Comparison of lidocaine,tramadol, and lidocaine–tramadol for epidural analgesia in lambs

Epidural anesthesia is commonly utilized in veterinary medicine to allow diagnostic, obstetrical, and surgical intervention, in the perineal region of domestic animal. The following study was carried out to directly compare the time of onset and duration of anesthesia produced by a tramadol and lidocaine–tramadol combination with that produced by lidocaine administration in the epidural space of lamb. Seven healthy female lambs of undefined breed weighing 15–20 kg were selected for this study. Epidural anesthesia was produced in all lambs by 2% lidocaine and with 2 weeks intervals repeated by combination of lidocaine–tramadol and tramadol alone. Analgesia was defined as lack of a response to pin prick test and pressure from hemostat clamp (closed to the first ratchet) applied first in the perineal area and then moved cranially toward the thoracic region until a response (movement associated with pin prick test or hemostat pressure) was observed. Time to onset, duration and cranial spread of analgesia were recorded. Heart rate (HR), respiratory rate (RR), and rectal temperature were recorded before (baseline, 0) and at 15, 30, 45, 60, 75, 90, 105 and 120 min after epidural administration of the solution. The results were expressed as mean ± SD and were analyzed by a one-way analysis of variance and Duncan’s test as a post hoc for heart rate, respiratory rate and body temperature and also, for time of onset and duration of analgesia. Graphpad Prism version 5 software program was used for all analyses. A value of P < 0.05 was considered significant. The tramadol produced a significant (P < 0.05) longer duration of analgesia than lidocaine alone and lidocaine–tramadol combination. Also, lidocaine–tramadol combination produced a significant (P < 0.05) longer duration of analgesia than lidocaine alone. Complete analgesia began more delayed in the tramadol treatment than lidocaine–tramadol and lidocaine alone. The combination of lidocaine–tramadol produced analgesia of longer duration than lidocaine and onset time was approximately same as lidocaine group.     

Panton–Valentine leukocidin and some exotoxins of <Emphasis Type="Italic">Staphylococcus aureus</Emphasis> and antimicrobial susceptibility profiles of staphylococci isolated from milks of small ruminants

The aims of this study were to determine the existence of pvl gene, some toxin genes, and mecA gene in Staphylococcus aureus strains isolated from sheep milk and to examine antimicrobial resistance profiles in staphylococci from sheep and goats' milk. The milk samples were collected from 13 different small ruminant farms in Kirikkale province from February to August 2009. A total of 1,604 half-udder milk samples from 857 ewes and 66 half-udder milk samples from 33 goats were collected. Staphylococcus spp. were isolated and identified from the samples. Toxin genes and mecA gene among S. aureus strains were determined by PCR. Antimicrobial susceptibility of staphylococci was examined by the disk diffusion method on Mueller-Hinton agar, and interpreted according to the Clinical Laboratory Standards Institute (CLSI) guidelines. The prevalence of subclinical intramammary infection in both ewes and goats was 5.2%. The most prevalent subclinical mastitis agents were coagulase-negative staphylococci and S. aureus with prevalences 2.8% (n:46) and 1.3% (n = 21), respectively. The prevalence of resistances in isolated Staphylococcus spp. to penicilin G, tetracycline, erythromycin, gentamicin, and enrofloxacin were found as 26.9% (18), 7.5% (5), 6.0% (4), 3.0% (2), and 1.5% (1), respectively. Only 3 of the 21 S. aureus ewe isolates (13.4%) were shown to harbor enterotoxin genes being either seh, sej or sec. However, fourteen (66.6%) of the 21 S. aureus isolates had pvl gene while none of the isolates harbored mecA gene. In conclusion, Staphylococci were shown to be the most prevalent bacteria isolated from subclinical mastitis of ewes and goats and these isolates were susceptible to most of the antibiotics. In addition, S. aureus strains isolated from ewes were harboring few staphylococcal enterotoxin genes. However, Panton–Valentine leukocidin produced by S. aureus could be an important virulence factor and contribute to subclinical mastitis pathogenicity.     

Cross-sectional survey of brucellosis and associated risk factors in the livestock–wildlife interface area of <Emphasis Type="Italic">Nechisar</Emphasis> National Park,Ethiopia

A cross-sectional survey was carried out to investigate the seroprevalence of ovine and bovine brucellosis in the livestock–wildlife interface area of Nechisar National Park, Ethiopia. Furthermore, producer’s knowledge about brucellosis and its zoonotic potential was assessed using a structured questionnaire. A total of 268 cattle and 246 goat sera were collected from 50 herds and 46 flocks and subjected to Rose Bengal test (RBT) and enzyme-linked immunosorbent assay (ELISA) in parallel to detect anti-Brucella species antibodies. Positive reactions were further confirmed with compliment fixation test (CFT). Flock and herd level seroprevalence rate was 12.8% (95% CI 4.8–25.7) and 32.0% (95% CI 19.5–46.7) in goats and cattle, respectively. An overall animal-level seroprevalence of 4.5% (95% CI 2.25–7.86) and 9.7% (95% CI 6.44–13.89) was recorded for goats and cattle, respectively. Seroprevalence showed an increasing trend with age, where adult cattle >?2 years. Goats (>?1 year) recorded relatively higher seroprevalence, but the differences were not statistically significant. Similarly, female cattle and goats recorded a relatively higher seroprevalence, 11 and 5.6%, respectively, compared to males but the difference was not significant. However, a significant (P <?0.01) variation of seroprevalence was noted for parity (bovine), higher in animals in second parity, and abortion history, in both species, higher in animals that experienced abortion. Interviews revealed lack of awareness about brucellosis and food safety related to the zoonotic potential from consuming raw animal products (milk and meat). Ninety-eight percent of respondents did not consider handling abortion material is risky, and only a very low proportion (8%, n?=?50) was able to mention limited zoonotic diseases (anthrax and Taenia cysticercosis) could be transmissible to people. The study indicated that brucellosis is endemic in domestic animals in the interface area and calls for further broad epidemiological investigation of the disease in livestock, human and wildlife following ‘one health’ unified research approaches beside enhancing public awareness.     

Reversible chemical restraint of free-range cattle with a concentrated combination of tiletamine–zolazepam,ketamine, and detomidine

The aim of this study was to determine the efficacy of a concentrated combination of tiletamine–zolazepam [TZ, 0.53 mg/kg body weight (BW)], ketamine (Ket, 0.53 mg/kg BW), and detomidine (Det, 0.04 mg/kg BW) in the immobilization of free-range cattle for clinical procedures. The combination was administered intramuscularly to 53 animals. Anesthesia was reversed with the α₂-adrenoceptor antagonist atipamezole. Locoregional anesthesia was provided with lidocaine when required. The TZKD combination induced suitable immobilization for minor surgical procedures or medical treatments. Anesthetic onset was rapid, taking a mean of 6.1 min [standard deviation (SD) 2.8 min]. The duration of anesthesia depended on the time of administration of the antagonist; the animals recovered in the standing position in 12.9 ± 8.9 min after the administration of atipamezole. The quality of anesthesia and analgesia were satisfactory. In conclusion, this TZKD combination can be used for both immobilization and minor surgical procedures in free-range cattle.     

Evaluation of sedative and antinociceptive effects of dexmedetomidine,midazolam and dexmedetomidine–midazolam in tegus (Salvator merianae)

Objective

To evaluate dexmedetomidine, midazolam and dexmedetomidine–midazolam for sedation and antinociception in tegus.

Serological status of Canadian cattle for brucellosis,anaplasmosis, and bluetongue in 2007–2008

A national bovine serological survey was conducted to confirm that the prevalence of brucellosis, bluetongue, and anaplasmosis does not exceed 0.02% (95% confidence) in live cattle in Canada. Sampling consisted of a systematic random sample of 15 482 adult cattle slaughtered in federally inspected abattoirs, stratified by province. Samples were tested to detect antibodies for brucellosis, bluetongue, and anaplasmosis. All samples were negative for brucellosis. Three samples were seroreactors to bluetongue, 2 of which originated from the Okanagan Valley in British Columbia and 1 from Ontario, which after follow-up, was considered an atypical result. A total of 244 samples were seroreactors to Anaplasma and follow-up identified infection in Saskatchewan, Manitoba, and Quebec. In conclusion, the Canadian cattle population remains free of brucellosis and free of bluetongue outside the Okanagan Valley. Canada is no longer free of anaplasmosis and will be unable to claim freedom until eradication measures are completed.