Viruses associated with outbreaks of equine respiratory disease in New Zealand

AIM: To identify viruses associated with respiratory disease in young horses in New Zealand.

METHODS: Nasal swabs and blood samples were collected from 45 foals or horses from five separate outbreaks of respiratory disease that occurred in New Zealand in 1996, and from 37 yearlings at the time of the annual yearling sales in January that same year. Virus isolation from nasal swabs and peripheral blood leukocytes (PBL) was undertaken and serum samples were tested for antibodies against equine herpesviruses (EHV-1, EHV-2, EHV-4 and EHV-5), equine rhinitis-A virus (ERAV), equine rhinitis-B virus (ERBV), equine adenovirus 1 (EAdV-1), equine arteritis virus (EAV), reovirus 3 and parainfluenza virus type 3 (PIV3).

RESULTS: Viruses were isolated from 24/94 (26%) nasal swab samples and from 77/80 (96%) PBL samples collected from both healthy horses and horses showing clinical signs of respiratory disease. All isolates were identified as EHV-2, EHV-4, EHV-5 or untyped EHV. Of the horses and foals tested, 59/82 (72%) were positive for EHV-1 and/or EHV-4 serum neutralising (SN) antibody on at least one sampling occasion, 52/82 (63%) for EHV-1-specific antibody tested by enzyme-linked immunosorbent assay (ELISA), 10/80 (13%) for ERAV SN antibody, 60/80 (75%) for ERBV SN antibody, and 42/80 (53%) for haemagglutination inhibition (HI) antibody to EAdV-1. None of the 64 serum samples tested were positive for antibodies to EAV, reovirus 3 or PIV3. Evidence of infection with all viruses tested was detected in both healthy horses and in horses showing clinical signs of respiratory disease. Recent EHV-2 infection was associated with the development of signs of respiratory disease among yearlings [relative risk (RR)=2.67, 95% CI=1.59-4.47, p=0.017].

CONCLUSIONS: Of the equine respiratory viruses detected in horses in New Zealand during this study, EHV-2 was most likely to be associated with respiratory disease. However, factors other than viral infection are probably important in the development of clinical signs of disease.     

The uterus and early pregnancy failure in the mare

The environment the equine conceptus finds itself in when it arrives in the uterus some 6 days after ovulation will determine if it will thrive or die. The uterus will be its home for approximately the next 11 months and as such it needs to both coordinate the growth of the placenta and ensure there is enough nourishment passing to this organ for the fetus to develop normally. The trophoblast, endometrium, maternal ovaries and, later in pregnancy, the fetal gonads, all play roles in the hormonal changes that orchestrate these events. Although failures of these processes later in pregnancy can have catastrophic effects for the fetus, it is in early gestation that the foundations for a successful pregnancy are laid. This paper therefore concentrates on some of the noninfectious influences the uterus may have on survival of the young conceptus.     

Poor performance associated with equine gastric ulceration syndrome in four Thoroughbred racehorses

Equine gastric ulceration syndrome (EGUS) is commonly recognised in Thoroughbred racehorses. Although EGUS has previously been associated with reduced athletic performance, no objective studies have been reported. This case report describes 4 racehorses referred for investigation of poor athletic performance where EGUS was the only abnormal finding during a thorough investigation of all body systems. All horses showed considerable improvement in performance following treatment with omeprazole. Therefore, this is the first report in which evidence is presented suggesting a direct link between EGUS and decreased performance, other causes of poor performance having been excluded.     

The range and prevalence of pathological abnormalities associated with lameness in working horses from developing countries

Reasons for performing study: Lameness is highly prevalent in working horses, but published reports on the associated pathological abnormalities are lacking. With over 42 million horses in developing countries and the majority used for work, lameness has major welfare implications. Objectives: To describe the range and prevalence of pathological abnormalities associated with lameness in working horses. Methods: A standard lameness assessment was adapted for field use in working equids. Data on pathological abnormalities and pain responses in the feet, limbs and spine were collected through observation, palpation, manipulations and gait assessment in working horses from India (n = 110) and Pakistan (n = 117). Lameness at the walk was scored on a scale of 0–4 (sound‐nonweightbearing). Results: All horses examined were lame. Overall, 98% showed a gait abnormality in all 4 limbs and 87% had at least one limb scoring 3 or 4 on the lameness scale. Multiple pathological abnormalities within each limb were associated with lameness, with similar results in both countries. Chronic foot pathology was seen in every horse; 94% horses showed signs of chronic joint disease; 83% had digital flexor tendonitis in at least one limb. Lameness and pathological abnormalities were associated with specific pain responses in the feet, limbs and spine. Conclusions: The extremely high prevalence of multilimb lameness and its association with pain is of great concern. The multiple pathological abnormalities present in working horses makes lameness complex to address. Potential relevance: The results of this detailed study of lameness should facilitate the identification of risk factors and the implementation of interventions to reduce the prevalence of lameness in working equids.     

Folliculogenesis, embryo parameters and post-transfer recipient pregnancy rate following equine follicle-stimulating hormone (eFSH) treatment in cycling donor mares

Background Induction of multiple ovulations, or superovulation, may potentially increase the efficiency of equine embryo transfer programs. Our objective was to investigate the effects of equine follicle‐stimulating hormone (eFSH) treatment on the success rate of embryo transfer programs in mares. Methods In the research facility of the University of Saskatchewan, Canada, we studied 12 donor mares and 37 recipient mares during the physiological breeding season. Donor mares were used in two consecutive oestrous cycles: the first served as the control cycle and in the second an eFSH regimen was applied (eFSH cycle). In the control cycle, mares were administered human chorionic gonadotropin (hCG) to induce ovulation when a follicle ≥35 mm in diameter was detected by transrectal ultrasonographic examination. In the second oestrous cycle, twice‐daily eFSH treatment was initiated when a follicle ≥25 mm was detected and treatment ceased when a follicle ≥35 mm was present, at which time hCG was administered. All donor mares were artificially inseminated while in oestrus using fresh semen collected from a stallion of proven fertility. At 8 days post‐ovulation, embryos were recovered transcervically and transferred individually to the uterus of a synchronised recipient mare. Results The eFSH treatment stimulated the ovary and resulted in greater numbers of ovulations and recovered embryos; however the recovered embryos tended to have a lower morphological grade than the control embryos, and the recipient pregnancy rate per transferred embryo was lower than anticipated. Conclusion The numbers of recipient pregnancies and foals born that resulted from eFSH treatment were not different from the control.     

Urinary incontinence and incoordination in three horses associated with equine protozoal myeloencephalitis

Urinary incontinence, weakness and ataxia associated with equine protozoal myeloencephalitis (EPM) was diagnosed in three horses. Rectal examination of all horses revealed distention of the urinary bladder. Urine was expressed when manual pressure was applied to the bladder of each horse during rectal examination. The anal reflex and tone of the anus and tail were normal in all horses. Two horses had bacterial cystitis associated with Enterococcus sp. All horses were treated with pyrimethamine and a sulfonamide for EPM, but there was a variable response to treatment.     

Morbidity and mortality associated with a Standardbred yearling sale

In the 2‐week period following a Standardbred yearling sale, 5 horses from the sale died or were subjected to euthanasia. Three were confirmed as equine herpesvirus type 1 myeloencephalopathy (EHM), while EHM was suspected in 2. Clinical abnormalities were reported in 77 of 177 (43.5%) other yearlings during the 14‐day period after the sale; however, no diagnostic testing was performed. Eleven secondary cases of suspected EHM in horses in contact with yearlings purchased at the sale were identified. No cases of EHM were identified associated with the same sale in 2003; however, there was no difference in morbidity with clinical abnormalities reported in 39/96 (41%) horses sold that year (P = 0.70). Yearling sales have an inherent risk of pathogen exposure and illness is common in purchased horses. Disease in these horses could have been associated with a wide range of respiratory pathogens; however, the incidence of confirmed or suspected primary and secondary EHM cases associated with the 2002 yearling sale highlight the potential for significant outbreaks of disease.     

Theiler's disease associated with administration of tetanus antitoxin contaminated with nonprimate (equine) hepacivirus and equine parvovirus-hepatitis virus

An 11-year-old Trakehner gelding was presented for evaluation of lethargy, decreased appetite, mild icterus, and elevated hepatic enzyme activities. Physical examination, serum chemistry results, and liver biopsy histopathologic findings were supportive of Theiler's disease. Polymerase chain reaction (PCR) testing results of serum and liver tissue were positive for nonprimate (equine) hepacivirus (NPHV) and a novel equine parvovirus-hepatitis virus (EqPV-H). PCR testing of the lot of tetanus antitoxin administered to the gelding 3 months previously also yielded positive results for NPHV and EqPV-H. Treatment included supportive care and clinical signs resolved within 1 week, although hepatic enzyme activities remained elevated for several months. The horse successfully returned to work as a hunter/jumper for about 1 year until it developed a forelimb lameness and progressive atrophy of shoulder musculature (sweeney), prompting a decision for euthanasia 20 months after initial evaluation. Serial PCR testing of serum revealed persistent infection with both NPHV and EqPV-H and necropsy examination revealed chronic active hepatitis, mild liver atrophy, and positive PCR results for NPHV and EqPV-H in liver tissue. This case highlights the possible risk of administering potentially contaminated biologics of equine origin and the importance of screening for recently identified hepatic viruses in donors from which blood products are prepared.     

Maternal periconceptual nutrition,early pregnancy,and developmental outcomes in beef cattle

The focus of this review is maternal nutrition during the periconceptual period and offspring developmental outcomes in beef cattle, with an emphasis on the first 50 d of gestation, which represents the embryonic period. Animal agriculture in general, and specifically the beef cattle industry, currently faces immense challenges. The world needs to significantly increase its output of animal food products by 2050 and beyond to meet the food security and agricultural sustainability needs of the rapidly growing human population. Consequently, efficient and sustainable approaches to livestock production are essential. Maternal nutritional status is a major factor that leads to developmental programming of offspring outcomes. Developmental programming refers to the influence of pre-and postnatal factors, such as inappropriate maternal nutrition, that affect growth and development and result in long-term consequences for health and productivity of the offspring. In this review, we discuss recent studies in which we and others have addressed the questions, “Is development programmed periconceptually?” and, if so, “Does it matter practically to the offspring in production settings?” The reviewed studies have demonstrated that the periconceptual period is important not only for pregnancy establishment but also may be a critical period during which fetal, placental, and potentially postnatal development and function are programmed. The evidence for fetal and placental programming during the periconceptual period is strong and implies that research efforts to mitigate the negative and foster the positive benefits of developmental programming need to include robust investigative efforts during the periconceptual period to better understand the implications for life-long health and productivity.     

Otitis media‐interna and secondary meningitis associated with Corynebacterium pseudotuberculosis infection in a horse

A one‐year‐old Thoroughbred colt was evaluated because of facial nerve paralysis, ataxia and fever. Neurological evaluation found the colt to be obtunded and grade 3/5 ataxic in all 4 limbs. Right‐sided facial nerve paralysis was present and a large, deep corneal ulcer noted in the right eye. Signs of vestibular disease were also present, including circling towards the right and horizontal nystagmus. A complete blood count showed mild leucocytosis, neutrophilia and hyperfibrinogenaemia. A computed tomography (CT) examination of the skull was performed under general anaesthesia and a diagnosis of right sided otitis media‐interna was made. Culture of fluid taken from the middle ear and cerebrospinal fluid collected from the atlanto‐occipital site yielded pure growth of Corynebacterium pseudotuberculosis. Initial therapy consisted of antimicrobial treatment with cefotaxime and anti‐inflammatory treatment with flunixin meglumine. Six days after initiating treatment, the colt developed Clostridium difficile associated colitis. The colitis resolved with supportive care and the colt was discharged from the hospital receiving chloramphenicol. Eight months later, the colt continued to be mildly ataxic (grade 1/5), with a slight head tilt and facial nerve paralysis. To the authors' knowledge, this is the first reported case of otitis media‐interna due to C. pseudotuberculosis in the horse.     

The effects of cooling and vitrification of embryos from mares treated with equine follicle-stimulating hormone on pregnancy rates after nonsurgical transfer

Equine embryos can remain viable for 12 to 24 hours when cooled and stored at 5°C.¹ Cryopreservation of embryos would allow for long-term preservation of genetic material and more efficient management of embryo recipients. This study compared pregnancy rates after transfer of equine embryos vitrified within 1 hour of collection or cooled for 12 to 19 hours before vitrification. Mares (N = 40) were superovulated using equine follicle-stimulating hormone (eFSH). Embryos were recovered 6.5 days after ovulation or 8 days after human chorionic gonadotropin. Forty morulae or early blastocysts with a grade of 1 to 2 and <300 mm in diameter were randomly assigned to 1 of 2 treatments: Group 1 (n = 20), washed 4 times in a commercial holding medium and then vitrified; Group 2 (n = 20), washed 3 times and then stored in the same holding medium at 5°C to 8°C in a passive cooling device for 12 to 19 hours before being vitrified. To thaw, embryos were warmed by holding the straw in air at room temperature for 10 seconds and then submerged in a water bath (20°C to 22°C) for an additional 10 seconds. The contents of the straw were transferred directly into a recipient that had ovulated 4 to 6 days previously. There were no differences (P > .05) in embryo diameter, grade, or morphology score between treatment groups before vitrification. Pregnancy rates (day 16) were not different (P > .05) between embryos vitrified immediately after collection (15 of 20; 75%) and embryos cooled for 12 to 19 hours before vitrification (13 of 20; 65%). Based on these results, small equine embryos (<300 mm) can be stored at 5°C to 8°C for 12 to 19 hours before vitrification without a significant loss of viability.     

Selection based on morphological features of porcine embryos produced by in vitro fertilization: Timing of early cleavages and the effect of polyspermy

The aim of this study was to examine whether a morphological approach is efficient for selecting high‐quality porcine embryos produced by in vitro fertilization (IVF) under high polyspermy conditions. Frozen‐thawed Meishan epididymal spermatozoa showing moderate and high polyspermy were subjected to IVF (1 × 10⁵ sperms/ml). Under conditions of moderate polyspermy, 4‐cell embryos selected at 48 hr after IVF (single selection) and 8‐cell embryos selected at 79 hr after IVF from the collected 4‐cell embryos (double selection) showed high developmental competence. Likewise, 4‐ and 8‐cell embryos produced by IVF under high polyspermy conditions also showed high competence for development to blastocysts. However, blastocysts derived from high polyspermy conditions had significantly fewer cells than those produced under moderate polyspermy conditions. Furthermore, the frequency of nuclear and chromosomal abnormalities in 4‐ and 8‐cell embryos produced under conditions of high polyspermy was significantly (p < .05) higher in comparison to moderate polyspermy conditions. These findings suggest that although high polyspermy affects the frequency of nuclear and chromosomal anomalies in porcine IVF embryos, subsequent selection based on morphological features of 4‐ and 8‐cell embryos even under high polyspermy conditions, could be an alternative option for selecting porcine IVF embryos with high development ability.     

Premature luteal regression in a pregnant mare and subsequent pregnancy maintenance with the use of oral altrenogest

Premature luteal demise or luteal insufficiency is not well characterised as a cause of pregnancy loss in domestic species, including horses. In this report, a mare inseminated with cooled‐transported semen at our facility returned for a routine pregnancy diagnosis at 15 days post ovulation. Ultrasonography per rectum revealed endometrial oedema and the absence of visual indication of a corpus luteum on either ovary. Nonetheless, an embryonic vesicle small for the gestational age was identified. Daily oral altrenogest treatment was implemented immediately. Serum progesterone concentration was 0.67 ng/ml, which is below the threshold considered adequate for pregnancy maintenance in the mare. Examinations were repeated at 17, 25, 30, 39, 49, 72 and 120 days post ovulation. At 25 days post ovulation the embryonic vesicle presented normal development for the gestational age. In addition, sequential blood samples were collected to measure progesterone, equine chorionic gonadotrophin and oestrone sulphate concentrations. Although progesterone concentration did not exceed 2 ng/ml until 72 days post ovulation, all other results were unremarkable and a healthy filly was born uneventfully at 344 days post ovulation.