Recent advances in understanding the epidemiology of Neospora caninum in cattle

INTRODUCTION: Neospora caninum is the most frequently diagnosed cause of cattle abortion in New Zealand and is an important pathogen worldwide. The parasite has been found in aborted bovine foetuses and in puppies with neurological disease. Recent developments have provided new insights into the epidemiology of the disease that are reviewed in this article. TRANSMISSION: Transplacental infection is of major importance in the spread of N. caninum overseas. Most congenital infections result in the birth of apparently healthy calves. Seroepidemiological studies indicate introduced point-source infections are also likely, as do investigations of abortion outbreaks in cattle herds attributed to N. caninum. Horizontal transmission is an important determinant of the stability of infection prevalence in a herd. Two potential sources of horizontal transmission that have recently been reported are: via a canine definitive host and; cow-to-calf transmission through pooled colostrum or milk. Transmission parameters for the organism have been estimated using mathematical modeling, to explore the epidemiology and options for control of N. caninum infection in dairy cattle in the absence of precise epidemiological data. LIFE-CYCLE: Seroepidemiologic studies, combined with the knowledge that dogs can be a definitive host, provide evidence supporting a dog-cattle life-cycle. The importance of dogs in the epidemiology of bovine neosporosis is not yet clear. It is likely that N. caninum oocysts in dog faeces could serve as a source of infection for cattle and recent studies have shown that the canine and bovine isolatesflare the same organism. Infection of cattle orally exposed to oocysts has been demonstrated experimentally but not in nature. It is not known if other wild carnivores may serve as definitive hosts of N. caninum. CONCLUSION: Despite active research for over a decade, very little is known about the mechanisms of transmission of N. caninum. Vertical transmission is well documented but horizontal spread and the pathogenesis of abortion from this disease need further investigation. It is evident that, even with a very high probability of vertical transmission, some form of horizontal transmission is required for the disease to persist in a herd.     

Ascarids. Recent advances

P. equorum is a common and ubiquitous parasite that persists for many years in stables and on pasture in spite of good hygiene and anthelmintic control programs. Foals are usually infected early in life. During the migratory phase of the infection, clinical signs include coughing and a nasal discharge followed by depression and unthriftiness as the worms mature in the gut. Some foals die as a result of intestinal impaction or rupture. Patency is established around 3 months of age, and fecal egg counts may rise to very high levels. From 6 months of age onwards, the ascarid burden diminishes as the foals become immune. Patent infections are seldom found in mature horses and, when present, they tend to be of low magnitude. Preventive measures are aimed at treating foals frequently enough to prevent the development of a large mass of ascarids in the intestine. This is achieved by a 6-weekly dosing regimen using an anthelmintic with proven and reliable efficacy against P. equorum.     

Extensive production of Neospora caninum tissue cysts in a carnivorous marsupial succumbing to experimental neosporosis

ABSTRACT: Experimental infections of Sminthopsis crassicaudata, the fat-tailed dunnart, a carnivorous marsupial widely distributed throughout the arid and semi-arid zones of Australia, show that this species can act as an intermediate host for Neospora caninum. In contrast to existing models that develop relatively few N. caninum tissue cysts, dunnarts offer a new animal model in which active neosporosis is dominated by tissue cyst production. The results provide evidence for a sylvatic life cycle of N. caninum in Australia between marsupials and wild dogs. It establishes the foundation for an investigation of the impact and costs of neosporosis to wildlife.     

Recent advances in avian virology.

Selected, recent research on the following avian diseases, and their causative viruses, has been reviewed: chicken anaemia, infectious bursal disease, turkey rhinotracheitis, avian nephritis, fowlpox, influenza, infectious bronchitis and turkey enteritis.     

Recent advances in inhalation anesthesia.

Both desflurane and sevoflurane offer theoretical and practical advantages over other inhalation anesthetics for horses. The lower solubility of both agents provides improved control of delivery and helps to counteract the confounding influence of the voluminous patient breathing circuit commonly used for anesthetizing horses. The lower solubility should account for faster rates of recovery compared with the older agents; whether or not the quality of recovery differs remains to be objectively evaluated in a broad range of circumstances. The pharmacodynamic effects are, in large part, similar to those of isoflurane (e.g., low arrhythmogenicity) but with some differences. For example, desflurane may be overall more sparing to cardiovascular function (especially during controlled ventilation) compared with isoflurane and sevoflurane, which are roughly similar. Respiratory depression with both new agents is equal to or more depressing than isoflurane, suggesting the use of mechanical ventilation, especially in circumstances of prolonged management (i.e., hours of anesthesia). Both new anesthetics, not surprisingly, are expensive. From this point there are some agent-unique considerations. The anesthetic potency of both agents is less than that of isoflurane, which influences the cost of anesthesia, but also places an upper limit on inspired oxygen concentration (of particular concern with desflurane). Both agents require new vaporizers, but because of the high boiling point and steep vapor-pressure curve of desflurane, new technology was required. This translates into more costly equipment, adding to the cost of desflurane use. In addition, electricity is necessary for the new desflurane vaporizer to function, which limits its portability and adds additional practical considerations in its clinical use. On the other hand, desflurane strongly resists degradation both in vitro and in vivo, but in vitro degradation of sevoflurane by CO2 absorbents may produce renal injury. This may be true especially in association with low fresh-gas inflow rates (used to reduce the cost of using the new agent), and university based practices, where prolonged anesthesia is common.     

Large strongyles. Recent advances

This review cites recent advances in the knowledge pertaining to infections of large strongyles in equids. Emphasis is placed on Strongylus vulgaris and attention is focused on pathogenesis of clinical manifestations of infections and treatment and control, including chemotherapy of migrating larvae and drug resistance.     

Small strongyles. Recent advances

The recent increased interest in cyathostomes can be traced to simplification of their taxonomy, improved knowledge of pathogenicity, and failures of practical control due to anthelmintic resistance. Cyathostome ova develop to infective third-stage larvae (L3) at a rate that is directly proportional to environmental temperature. Equine feces serve as a reservoir for L3, which are liberated by moderate amounts of rainfall. Third-stage larvae persist for longer periods at low temperatures, easily surviving over-winter on pastures to provide a source of infection during the following grazing season. Third-stage larvae exsheath within the host and enter the mucosa and submucosa of the cecum and large colon. Larvae develop within mucosal cysts, molt to the fourth stage, and may persist within the tissues for up to 2 1/2 years. Larvae ultimately emerge from the mucosa to become adults in the lumen. Adult populations are replenished by recently ingested larvae and by immature worms newly emerged from arrested development. The magnitude of larval and adult populations within the host displays seasonal variations, with peak numbers occurring in early spring and autumn in the United States. In typical natural infections, a small number of species comprise the majority of the cyathostome populations. Cyathostome infection may result in anorexia, weight loss, diarrhea, colic, and death. Cyathostome ova are easily detected in feces, but ova may not be present during larval cyathostomiasis. Increased concentrations of beta-globulins, hypoalbuminemia, anemia, and leukocytosis occur inconsistently. Two major problems in the treatment of cyathostome infections are anthelmintic resistance and the insusceptibility of encysted larvae to recommended dosages of most anthelmintics. The major goal of cyathostome control is prevention of environmental contamination with nematode ova. Host resistance appears to protect against cyathostome disease rather than cyathostome infection, and one manifestation of this resistance appears to be prolongation of the prepatent period.     

Recent advances in small animal genetics.

The whole genome sequence of the dog is complete, and partial sequencing of the cat genome is underway. Sequences allow the molecular basis for inherited diseases to be more easily determined, leading to development of DNA tests to verify carrier and affected states as well as potential gene therapy for the treatment of those diseases. To help veterinarians provide genetic services to their clients, the molecular genetic tests currently available are listed in this article. In addition, cloning of small animals is now available to clients on a commercial basis. Information about the cloning process and possible health issues in clones are discussed.     

Artificial insemination of cows with semen in vitro contaminated with Neospora caninum tachyzoites failed to induce neosporosis

Neosporosis is a major cause of abortion in cattle all over the world. Congenital transmission as well as horizontal transmission by ingestion of oocysts has been described. The detection of Neospora caninum DNA in bull semen warrants the investigation of possible transmission through the use of contaminated semen. In this experiment four cows were artificially inseminated with frozen-thawed semen contaminated in vitro with viable N. caninum tachyzoites (group A) and four control cows were inseminated with tachyzoites-free frozen-thawed semen, from the same bull (group B). Serum samples were collected 15 days before the artificial insemination (AI) and at days 10, 14, 21, 28, 45, 60 and 75 post-insemination. All sera samples were tested for neosporosis by direct agglutination test (DAT). Three of the cows from group A had negative DAT titers (< or =1:20) in all of the samples, while the fourth cow from this group had a low titer of antibodies (1:80) at day 10, and became negative at day 45, suggesting a stimulation of the immune system by the tachyzoites placed in uterus, rather than the induction of an infection. All of the cows from group B had negative DAT titers (< or =1:20) in all of the samples. These results suggest that transmission of neosporosis by artificial insemination with frozen-thawed semen is an unlikely event.     

Recent advances in respiratory conditions in cattle.

The study of cattle respiratory diseases during the last decade show that several observations made in the late 1960s are still relevant today. Many of these diseases are shared with other countries in Europe, but the developments discussed will be examined with the respiratory disease problems that occur in Britain in mind. The new information is dealth with as it applies to (i) structure, function and immunology of the lungs (ii) diseases of calves indoors (iii) diseases of young animals grazing, (iv) diseases of adult animals indoors and (v) diseases of adult animals grazing.     

Canine neosporosis in South Africa.

Neospora caninum was identified in tissues of one neonatal and two adolescent dogs by a specific immunohistochemical technique during a retrospective study. The histological lesions included multifocal polymyositis, multifocal parasitic myocarditis, interstitial pneumonia, severe multifocal encephalitis and myelitis with perivascular cuffing. This report extends the known range of the parasite to the African continent.     

Evaluation of the protection conferred by a naturally attenuated Neospora caninum isolate against congenital and cerebral neosporosis in mice

The parasite Neospora caninum is an important abortifacient agent in cattle worldwide. At present, the development of an effective and safe vaccine against bovine neosporosis is of great relevance. Recently, a new isolate of N. caninum (Nc-Spain 1 H) which was obtained from the brain of an asymptomatic congenitally infected calf, exhibited non-virulent behaviour in mouse and bovine infection models. The aim of this study was to determine the safety and efficacy of Nc-Spain 1 H when used as a vaccinal isolate in well-established BALB/c models of congenital and cerebral neosporosis. Mice were subcutaneously immunised twice at 3-week intervals and were challenged with 2 × 10⁶ tachyzoites of the virulent Nc-Liv isolate. After immunisation with live Nc-Spain 1 H tachyzoites, no parasitic DNA was detected in the dams’ brains before challenge and microsatellite analysis performed in PCR-positive mice showed that the profiles corresponded to the challenge isolate Nc-Liv, indicating the Nc-Spain 1 H isolate to be a safe vaccine candidate. The efficacy of the live vaccine was evaluated in the first experiment after the immunisation of mice with 5 × 10⁵ live Nc-Spain 1 H tachyzoites. This immunisation protocol significantly reduced the neonatal mortality to 2.4%, reduced the vertical transmission from 89.1% to 2.3% and completely limited the cerebral infection. These results were associated with a Th1-type immune response. In the second experiment, the effect of various immunising doses was established using ten-fold dilutions of the tachyzoites (from 5 × 10⁵ to 5 × 10). In all the cases, congenital protection rates above 60% were observed, and the mice that were immunised with the lowest dose (5 × 10) presented the highest protection rate (86%). Moreover, low immunising doses of Nc-Spain 1 H induced an IgG2a response, and high parasitic doses induced an IgG1 response. These results evidence the safety and the efficient protection that was conferred by Nc-Spain 1 H against congenital neosporosis, even when the mice were immunised with low parasitic doses.     

Distribution of Neospora caninum within the central nervous system and other tissues of six dogs with clinical neosporosis

Six cases of neosporosis in dogs were diagnosed on the basis of clinical signs, serology and immunohistochemistry. The brains and spinal cords, at several levels, and various other tissues were examined using anti-Neospora caninum serum by an immunoperoxidase technique. Parasite density varied markedly from case to case. Although found most consistently in the cerebrum, parasites were distributed throughout the central nervous system (CNS), both within the grey and white matter and within nerve roots. Clinical signs were not related to the position of parasites. Tissue cysts were found infrequently in all areas of the CNS, but not in other tissues. Parasite density was not related to the age of the dog or whether treatment had been given. However, with the exception of the only adult dog examined, more CNS parasites were found in dogs with a longer duration of illness and with higher antibody titres. Tachyzoites were present in skeletal muscles, in the muscularis of the oesophagus, in heart, lung and, less frequently, liver, and rarely in the adrenal gland, thyroid gland and uterus; no clinical signs were seen resulting from damage to these organs. Parasites were not observed in lymphoid tissue. In visceral organs, parasites were most widely distributed in peracute cases. For post mortem diagnosis of neosporosis, the CNS, particularly the cerebrum, is the optimum tissue to examine but parasites may also be found in many other tissues, especially in acute cases. Muscle biopsy of appropriate muscles (as suggested by the clinical signs) provides the possibility of a definitive pre-mortem diagnosis.     

First Portuguese isolate of Neospora caninum from an aborted fetus from a dairy herd with endemic neosporosis

Neospora caninum was isolated from the brain of an aborted 4-month-old fetus from a dairy cow herd with endemic neosporosis in Porto, Portugal. The fetal brain homogenate was inoculated interperitoneally first into outbred Swiss Webster mice given dexamethasone and then the peritoneal exudates from these mice was co-inoculated with mouse sarcoma cells in the peritoneal cavity of mice given dexamethasone. N. caninum tachyzoites were seen in peritoneal exudate of the second passage. Tachyzoites from the peritoneal exudate reacted positively with anti-N. caninum antibodies and not with anti-Toxoplasma gondii antibodies and contained N. caninum specific DNA. This Portuguese isolate of N. caninum has been successfully maintained in cell culture. The dam of the aborted fetus had an antibody titer of 1:10240 in the Neospora agglutination test (NAT). Antibodies to N. caninum were found in 76 of 106 cows from this herd in titers of 1:40 in 31, 1:80 in 22, > or =1:160 or more in 23 in the Neospora agglutination test. This is the first isolation of a viable N. caninum-like parasite from any host in Portugal.     

Recent advances in the field of doping detection.

Patterns of doping in Britain have on the whole changed from the use of CNS stimulants and depressants to therapeutic substances, or to accidental doping by feeding contaminated food or food supplements. Progress in detection and identification is largely a result of improvements in separation and isolation, especially by gas chromatography, together with the introduction of mass spectrometry. Radiommunoassay and high performance liquid chromatography promise to be important techniques in the future, especially in detection of anabolic steroids and corticosteroids.     

Recent advances in the treatment of vaccine-associated sarcomas.

This article reviews the background information about vaccine-associated sarcomas followed by diagnostic procedures essential to understand how to determine the extent of the primary and metastatic tumor as well as to understand the general health of the patient. It also addresses the importance of understanding the nonmedical needs of the client who is faced with this perplexing problem.     

Evaluation of vaccination with Neospora caninum protein for prevention of fetal loss associated with experimentally induced neosporosis in sheep

OBJECTIVE: To evaluate the immunologic response of a killed tachyzoite vaccine against Neospora caninum and its effectiveness in preventing fetal loss associated with experimentally induced neosporosis in sheep. ANIMALS: 30 Dorset ewes. PROCEDURE: Ewes were randomly allocated to receive vaccination on days 1 and 60 of the study with a killed N caninum tachyzoite preparation in a commercially available adjuvant or a saline-adjuvant mixture. A ram was placed on pasture with the ewes from days 15 to 60. Blood was collected from ewes before primary and booster vaccinations and prior to experimental challenge with N caninum tachyzoite performed on day 90; sera were assessed via Neospora agglutination (NA) and immunofluorescence antibody (IFA) assays. Blood was collected from lambs before they suckled, and sera were tested for antibodies against N caninum. RESULTS: Of the 14 vaccinated ewes that became pregnant, 12 gave birth to live-born lambs; in contrast, 5 of 11 pregnant control ewes gave birth to live-born lambs. Whereas vaccination improved fetal survival in pregnant ewes challenged with N caninum tachyzoites, it did not appear to have any appreciable effect on transmission of N caninum to offspring, as indicated by results of NA and IFA assays. CONCLUSIONS AND CLINICAL RELEVANCE: The N caninum tachyzoite vaccine used in this study appeared to provide protection against fetal loss associated with experimentally induced neosporosis in a high proportion of pregnant ewes.