Epidemiology of leptospirosis in dairy farm workers in the Manawatu. Part II. A case-control study of high and low risk farms

Subsequent to a cross-sectional serological survey of Manawatu dairy farm workers, a case-control study was carried out to investigate the correlation between titres to leptospiral serovars in workers and those in cattle in their herds. A total of 52 herds was investigated, 25 of which were ‘high risk’ where milkers had titres of 1:96 or greater, and 27 were case-controls where milkers had no detectable agglutinin titres at a minimum serum dilution of 1:24.

The serological prevalence of titres to hardjo in cattle on ‘high risk’ farms (76.5%) was significantly higher (P<0.05) than on the case-control farms (60.0%). The geometric mean titres of seropositive cattle on ‘high risk’ farms were also significantly higher (P<0.0l) than in the cattle from the case-control farms, especially in the younger cohorts. These findings suggest that there was active endemic hardjo infection in the two- to three-year-old cattle on the ‘high risk’ farms.

Titres to pomona were demonstrated in only 5.2% of the cattle from both types of farm. Workers with titres to pomona tended to be from farms on which stock, especially calves, were bought-in and pigs were kept.

Conventional measures for protecting milkers from contact with infected urine appeared to be ineffective and it is concluded that prevention of leptospirosis in dairy farm workers can only be achieved by elimination of infection in the herd by vaccination of cattle.     

Investigation of bovine haemoplasmas and their association with anaemia in New Zealand cattle

CASE HISTORY AND CLINICAL FINDINGS: A dairy cow, from a herd in the Waikato region of New Zealand, was reported with regenerative anaemia on 12 September 2014. Testing of blood from the animal using PCR assays for Theileria orientalis produced a negative result for both Chitose and Ikeda types.

LABORATORY FINDINGS: Using PCR and DNA sequencing, blood from the cow was positive for Candidatus Mycoplasma haemobos. Further testing of another 12 animals from the case herd, 27 days after the affected cow was first reported, showed 11 animals were positive for Candidatus M. haemobos or Mycoplasma wenyonii in the PCR. None of these cattle were clinically anaemic or positive for T. orientalis Ikeda type using PCR.

A convenience sample of 47 blood samples from cattle throughout New Zealand, submitted to the Investigation and Diagnostic Centre (Ministry for Primary Industries) for surveillance testing for T. orientalis Ikeda, was selected for further testing for bovine haemoplasmas. Of these samples, 6/47 (13%) and 13/47(28%) were positive for M. wenyonii and Candidatus M. haemobos, respectively. There was no difference in the proportion of samples positive for the bovine haemaplasmas between cattle with anaemia that were negative for T. orientalis (6/20, 33%), or without anaemia or T. orientalis (10/18, 56%), or from cattle herds experiencing anaemia and infection with T. orientalis Ikeda type (3/9, 33%).

DIAGNOSIS: Bovine haemoplasmosis.

CLINICAL RELEVANCE: The presence of bovine haemoplasmas in blood does not establish causality for anaemia in cattle. Diagnosis of anaemia associated with haemoplasmosis would require exclusion of other causes of regenerative anaemia and an association of the agent with anaemia in affected cattle herds. The data collected in this study did not provide evidence that bovine haemoplasmas were associated with a large number of outbreaks of anaemia in cattle in New Zealand.     

Nasal infection with Scedosporium apiospermum in a dog

CASE HISTORY: A 2-year-old female Siberian Husky was presented with a 6-month history of sneezing and mucous discharge from the right nostril.

CLINICAL FINDINGS: Reduced airflow through the right nostril was evident. Radiographs showed subtle loss of detail of turbinates within the right nasal chamber. Rhinoscopy revealed swollen and erythematous turbinates and a white mass within the caudal aspect of the right nasal cavity. Histopathologically, there was a heavy mixed inflammatory infiltrate in the submus- cosa of the right turbinate, and the presence of fungal hyphae and spores in the white mass. A heavy growth of Scedosporium apiospermum was cultured from the mass.

DIAGNOSIS: Chronic rhinitis of the right nasal cavity and infection with S. apiospermum.

CLINICAL RELEVANCE: This is the first reported case of S. apiospermum isolated from the nasal cavity of a dog in New Zealand. Fungal culture is necessary to differentiate this fungus from Aspergillus spp.     

Investigation of the index case herd and identification of the genotypes of Theileria orientalis associated with outbreaks of bovine anaemia in New Zealand in 2012

CASE HISTORY AND CLINICAL FINDINGS: On 7 September 2012 the Ministry for Primary Industries was notified of a dairy cow with regenerative anaemia (haematocrit (HCT) 0.08?L/L) in a herd of 465 Jersey-Friesian cross cows (index case herd) in the Northland region of New Zealand. Organisms consistent with Theileria spp. were present in red blood cells on a blood smear. No other causes of anaemia were detected following examination of affected cows. Blood samples collected from 29 randomly selected cows on 26 September 2012 showed that 24 (83%) were anaemic (HCT≤0.24 L/L) and therefore fitted the case definition for bovine anaemia associated with Theileria orientalis infection.

LABORATORY FINDINGS: Using a T. orientalis type-specific PCR assay that targeted the single subunit rRNA gene, all of six animals tested were positive for T. orientalis type Ikeda. Blood samples collected from clinically affected cattle in 11 subsequent outbreaks from throughout the North Island showed that T. orientalis Ikeda type was a common finding, but mixed infections with Chitose type were also identified. In addition, using a PCR assay that targeted the major piroplasm surface gene, T. orientalis type 5 was detected in one cow from the Waikato region.

DIAGNOSIS: The presence of T. orientalis type Ikeda, as well as type 5, was confirmed in cattle from outbreaks of bovine anaemia in herds throughout the North Island of New Zealand.

CLINICAL RELEVANCE: Two new types of T. orientalis were identified in this investigation, that were associated with a sudden rise in cases of bovine anaemia. The body of evidence showed that the Ikeda type was implicated as the cause of disease observed in this epidemic.     

Descriptive summary of an outbreak of porcine post-weaning multisystemic wasting syndrome (PMWS) in New Zealand

CASE HISTORY: Investigations were conducted to determine the cause of an acute, multi-farm outbreak of porcine respiratory disease that included diarrhoea and subsequent loss of body condition in affected pigs. A definition for post-weaning multisystemic wasting syndrome (PMWS) including both clinical and pathological features, previously developed for the pig industry in New Zealand, was applied to the current outbreak. In addition to self-reporting by owners of affected farms, local veterinarians, disease and epidemiology consultants, and animal health officials from the Ministry of Agriculture and Forestry (MAF) were involved in conducting farm visits and submission of diagnostic specimens.

CLINICAL FINDINGS AND DIAGNOSIS: Pathogens known to be endemic in the pig industry in New Zealand as well as likely exotic diseases were excluded as causative agents of the outbreak. Clinical signs including dyspnoea, diarrhoea, and rapid loss of body condition were consistent with the New Zealand case definition for PMWS. Interstitial pneumonia, pulmonary oedema, generalised lymph-node enlargement, and presence of porcine circovirus type 2 (PCV2) inclusion bodies were consistently identified in affected pigs. Classical swine fever virus (CSFv), Porcine reproductive and respiratory syndrome virus (PRRSv), and Influenza virus were ruled out, using molecular and traditional virological techniques. Spread of the disease between farms was hypothesised to be facilitated by locally migrating flocks of black-backed seagulls. The original source of the disease incursion was not identified.

DIAGNOSIS: Based on the consistent presence of circovirusassociated lesions in lymphoid tissues in combination with generalised enlargement of lymph nodes, histiocytic interstitial pneumonia, clinical wasting, and poor response to antibiotic therapy, a diagnosis of PMWS was made.

CLINICAL RELEVANCE: PMWS should be considered in the differential diagnoses of sudden onset of respiratory dyspnoea, diarrhoea, and rapid loss of body condition in young pigs in New Zealand pig herds.     

A serological survey of cattle in the Thames-Coromandel district of New Zealand for antibodies to Ross River virus

AIM: To determine if cattle exposed to the southern saltmarsh mosquito (SSM), Aedes camptorhynchus, in the Thames-Coromandel district of New Zealand had been exposed to Ross River virus (RRV).

METHODS: A purposive sampling design was used to test cattle from seven farms located in close proximity to four sites infested with A. camptorhynchus in the Thames-Coromandel district. Sera from 207 cattle were tested for antibodies to RRV, using an ELISA and confi rmatory virus neutralisation test (VNT) as the gold standard.

RESULTS: All 207 cattle tested negative for antibodies to RRV using the ELISA and VNT.

CONCLUSIONS: This study found no evidence of exposure to RRV in cattle in locations in the Thames-Coromandel district of New Zealand where populations of SSM were present.     

Prevalence of antibodies to Neospora caninum in dogs of rural or urban origin in central New Zealand

AIM: To investigate the seroprevalence of Neospora caninum infection in populations of dogs from dairy farms, sheep/beef farms and urban areas in the central part of New Zealand. It was postulated seroprevalence would be higher for farm dogs than urban dogs if the life-cycle of this parasite involves transmission between dogs and cattle.

METHODS: Serum samples were obtained from dogs that lived on dairy farms (n=161), sheep/beef farms (n=154) and in urban situations (n=150). The relative risk of detecting antibodies to N. caninum using an immunofluorescent antibody test (IFAT) was compared between farm and urban dogs.

RESULTS: The relative risk of having a titre of ≥1:200 to N. caninum was 2.43 (95% CI=1.88-3.14) for dairy-farm dogs and 3.16 (95% CI=2.48–4.02) for sheep/beef-farm dogs, compared with urban dogs. At this titre, which is currently used in New Zealand to indicate seropositivity, seroprevalence of N. caninum infection was 30.7% in urban dogs, 74.5% in dairy-farm dogs and 96.8% in sheep/beef-farm dogs.

CONCLUSION: This observation is consistent with a cycling of this disease between cattle and dogs on farms in New Zealand and with higher exposure of dogs to N. caninum on farms than occurs in urban environments. The prevalence of antibodies in all three groups of dogs tested in this study (dairy-farm dogs, sheep/beef-farm dogs and urban dogs) is higher than has generally been reported elsewhere. New Zealand farm dogs have a higher serological prevalence of N. caninum infection than urban dogs.

CLINICAL RELEVANCE: Management and disease control practices that break the life-cycle of transmission between cattle and dogs should assist in controlling cattle abortion due to N. caninum.     

A syndrome of facial paralysis of dairy calves in the Franklin district of New Zealand

AIM: To determine the aetiology of a syndrome characterised by facial paralysis in calves (facial paralysis syndrome; FPS); describe the epidemiology of the syndrome on an affected case farm; and define the intra-farm prevalence of affected calves, and inter-farm prevalence of affected dairy farms, in the Franklin district of New Zealand.

CASE HISTORY AND CLINICAL FINDINGS: An investigation was carried out on a town-supply dairy farm experiencing an outbreak of FPS in calves during the autumn of 2007, following a previous outbreak during the spring of 2006; 21 calves were affected in both outbreaks. Post-mortem examinations of three affected calves revealed no infectious aetiological agent in neurological tissues despite tests for viruses, bacteria and Mycoplasma species. Tests on hepatic tissues for vanadium toxicity were inconclusive.

SURVEY OF DAIRY FARMS: Results from a postal survey of 177/325 (54%) farms established the yearly prevalence of affected farms, based on farmer diagnosis, was 11%, and there was a median two (range 1–25) affected calves on those farms. There was no evidence of spatial clustering of affected farms after accounting for the underlying farm density, or of an increase in the number of affected farms between 2003 and 2007.

CLINICAL RELEVANCE: Facial paralysis syndrome is an unusual condition that has not been reported in other districts of New Zealand or in other countries. It is probable that this syndrome will continue to occur at a low to moderate prevalence, and have a significant impact on a small number of farms.     

Induction of diplodiosis, a neuromycotoxicosis, in domestic ruminants with cultures of indigenous and exotic isolates of Diplodia maydis

Diplodiosis, a neuromycotoxicosis, principally of cattle, which is characterized by ataxia, paresis and paralysis, was induced in 13 cattle, 16 sheep and 3 goats, by dosing them with Diplodia maydis [= D. zeae (Schw.) Lév.] cultured on sterilized maize seeds. The results of these experiments confirmed the findings of earlier workers that diplodiosis is a mycotoxicosis caused by D. maydis. The intoxication was induced with cultures of South African isolates of D. maydis obtained from local maize, one of which was associated with a suspected field outbreak, and with cultures of isolates from maize imported from the United States of America and Argentina. Other findings emerging from the experiments were, inter alia, that cultures incubated for less than 8 weeks were seemingly non-toxic, that there was little individual variation in response of cattle to cultures of the different toxic isolates or batches of the isolates, that apparent relapses of clinical signs can occur several weeks after dosing had ceased and that a small percentage of animals can show permanent locomotory disturbance. Light microscopical examination revealed no lesions in acutely affected animals, but an extensive laminar subcortical status spongiosis was evident in the cerebrum and cerebellum of a sheep that had been long paralysed and a steer that had permanent locomotory disturbance.     

Case Report: Sequelae of traumatic reticuloperitonitis in a Friesian dairy cow

Abstract

CASE HISTORY: A 10-year-old Friesian dairy cow presented with a history of weight loss, decreased milk production and fluid splashing on auscultation of the heart. Previous antibiotic therapy included oxytetracycline, marbofloxacin and penicillin.

CLINICAL AND PATHOLOGICAL FINDINGS: Following a failure to respond to treatment, prescribed by the referring veterinarian for respiratory infection and septic arthritis, the cow was referred to Massey University Veterinary Teaching Hospital with suspected pericarditis. Clinical examination identified a predominantly left sided holodiastolic heart murmur with tachycardia and cording of the jugular veins. Cardiac ultrasound showed an enlarged aortic valve with turbulent blood flow and regurgitation, consistent with bacterial endocarditis. Blood culture demonstrated Paenibacillus spp. Due to the severity of the clinical signs and poor prognosis, euthanasia was performed and a post-mortem examination carried out.

DIAGNOSIS: Traumatic reticuloperitonitis with abscessation of the reticulum and rumen wall, which progressed to a bacteraemia and presumptively caused endocarditis, endometritis, septic arthritis, and renal and myocardial infarcts due to emboli from the endocarditis lesions.

CLINICAL RELEVANCE: New Zealand dairy cattle are at risk of traumatic reticuloperitonitis and due to varying presentations diagnosis can be complicated. Subsequent bacteraemia is common and in this case the novel bacterial species Paenibacillus was implicated as a potential pathogen.     

A comparison of highveld fodders and fodder flows for growing out long yearling steers

Abstract

A fodder flow/animal experiment 40,8 ha in extent (7,5 ha irrigated), with 19 paddocks, was carried out over a two and a half year period.

The aims were: firstly, to compare fodder crops within seasons, and fodder flows over three seasons (winter‐spring‐summer), by means of pasture and animal parameters; secondly, to make economic comparisons between different fodder flows, for growing out beef steers.

The conclusions were that: Digitaria eriantha ssp. eriantha is superior to Eragrostis curvula cv. Ermelo for animal production; maize residue gives economically favourable results due to low costs and compensatory growth of cattle; an irrigated pasture providing fodder in more than one season of a flow, for example, Festuca foggage — Festuca pasture — Digitaria pasture, gives a very profitable fodder flow, because the costs of production are divided between two seasons; dryland fodder flows with the sequence maize residue — silage or hay — Digitaria or, with silage or hay replacing the maize residue, were also very profitable; and, the fodder flow giving the highest livemass gain per steer, is not necessarily the most economic. Different kinds and classes of livestock require different fodder flows.     

Necrotising fasciitis in a bull due to infection with Arcanobacterium haemolyticum

Abstract

CASE HISTORY: A 2-year-old Hereford bull was lame for one week before becoming recumbent.

CLINICAL AND PATHOLOGICAL FINDINGS: The scrotum and ventral perineal region were cold and blackened caudally. The semimembranosus and semitendinosus muscles were firm on palpation. The bull was subject to euthanasia, and necropsy revealed that the skin and S/C tissues of the caudal half of the scrotum were grey and necrotic. The caudal and distal aspects of the semimembranosus and semitendinosus muscles were grey and necrotic to a depth of approximately 15 cm, and these changes appeared to track along fascial planes. The tissue had an offensive smell, and large amounts of flocculent, watery, brown fluid and some gas were present. Histology of affected muscle and S/C tissues revealed coagulative necrosis, with oedema and large numbers of bacteria that were predominantly Gram-positive rods. Adjacent blood vessels contained thrombi while the epidermis overlying the affected areas appeared diffusely necrotic, suggesting infarction. Culture of the fluid yielded a pure growth of Arcanobacterium spp., which was identified as Arcanobacterium haemolyticum, using an API Coryne biochemical test strip.

DIAGNOSIS: Necrotising fasciitis and myositis due to Arcanobacterium haemolyticum.

CLINICAL RELEVANCE: Arcanobacterium haemolyticum has not previously been reported as a cause of necrotising fasciitis in any species. Necrotising fasciitis is probably an under-reported condition in cattle due to its clinical similarity to clostridial disease.     

An outbreak of toe ulcers,sole ulcers and white line disease in a group of dairy heifers immediately after calving

Abstract

CASE HISTORY: An outbreak of severe lameness was reported on a 780-cow spring-calving dairy herd in the Manawatu region within 1–3 weeks of calving, which affected over 90% of a group of 150 dairy heifers. Approximately 3 weeks before the planned start of calving heifers had been fed maize and grass silage on a concrete feed pad for 3–4 h per day, and mixed with a group of adult cows.

CLINICAL FINDINGS: During treatment for lameness, the feet of 28 lame heifers were closely examined. Seventy lesions were recorded in the claws of these heifers. The predominant lesion was severe solar and white line haemorrhage, with much of the haemorrhage focussed at the site at which sole ulcers are normally seen. White line disease was seen in nine claws and sole ulcers in four. Additionally, a lesion which is not commonly recorded in New Zealand, a toe abscess arising from separation of the white line from the sole in the abaxial region of tip of the toe was recorded in 11 claws. The clinical impression was that the soles of the affected heifers were very thin. This was supported by examination of the feet of eight heifers using ultrasonography that indicated heifers with hoof horn haemorrhages had very thin soles (estimated mean 4.1 mm) and that these soles were thinner than those in non-lame heifers without haemorrhages.

DIAGNOSIS: The pattern of disease seen in these heifers closely matched that seen in an outbreak of lameness in heifers in Florida, which was linked to thin soles resulting from excess hoof horn wear. The clinical signs and findings of the examination using ultrasonography strongly implicated thin soles as the underlying cause of lameness on this farm. An on-farm investigation revealed a combination of heifer behaviour and prolonged exposure to wet concrete on the feed pad were the primary causes of excess wear that resulted in thin soles.

CLINICAL RELEVANCE: This was an unusual outbreak of lameness in a group of newly calved heifers. The problem highlights the value of good management during the transition of heifers at pasture to the lactating herd standing on concrete for long periods, in the control of lameness.     

A tremorgenic mycotoxicosis of cattle caused by maize sprouts infested with Aspergillus clavatus

An outbreak of disease affecting a herd of 16 dairy cattle which were fed mouldy, sprouted maize is described. Eight of the cattle were affected, 5 of which died. The clinical signs included muscular tremors, hypersensitivity, ataxia, anorexia and salivation. Aspergillus clavatus was the only fungus isolated from the sprouts. Clinical signs that were indistinguishable from those in the field outbreak were reproduced by dosing the mouldy maize sprouts to a steer and a sheep, and by dosing another sheep with maize inoculated with a pure culture of A. clavatus isolated from the mouldy maize on the farm. Light microscopical examination revealed neuronal degeneration and necrosis in the midbrain, medulla oblongata and spinal cord of all 3 of these animals. The disease is clinically and pathologically indistinguishable from the disease caused by the ingestion of sorghum beer residue, and in certain respects it is similar to toxicoses caused by the ingestion of wheat sprouts and malt sprouts infested with A. clavatus.     

Clinical haematology and biochemistry profiles of cattle naturally infected with Theileria orientalis Ikeda type in New Zealand

AIMS: To present the haematology and biochemistry profiles for cattle in New Zealand naturally infected with Theileria orientalis Ikeda type and investigate if the results differed between adult dairy cattle and calves aged <6 months.

METHODS: Haematology and biochemistry results were obtained from blood samples from cattle which tested positive for T. orientalis Ikeda type by PCR, that were submitted to veterinary laboratories in New Zealand between October 2012 and November 2014. Data sets for haematology and biochemistry results were prepared for adult dairy cattle (n=62 and 28, respectively) and calves aged <6 months (n=62 and 28, respectively), which were matched on the basis of individual haematocrit (HCT). Results were compared between age groups when categorised by HCT. Selected variables were plotted against individual HCT, and locally weighted scatterplot smoothing (Loess) curves were fitted to the data for adult dairy cattle and calves <6 months old.

RESULTS: When categorised by HCT, the proportion of samples with HCT <0.15 L/L (severe anaemia) was greater for adult dairy cattle than for beef or dairy calves, for both haematology (p<0.002) and biochemistry (p<0.001) submissions. There were differences (p<0.05) between adult dairy cattle and calves aged <6 months in the relationships between HCT and red blood cell counts, mean corpuscular volume, mean corpuscular haemoglobin, mean corpuscular haemoglobin concentrations, lymphocyte and eosinophil counts, and activities of glutamate dehydrogenase and aspartate aminotransferase. In both age groups anisocytosis was frequently recorded. The proportion of blood smears showing mild and moderate macrocytosis was greater in adults than calves (p=0.01), and mild and moderate poikilocytosis was greater in calves than adults (p=0.005).

CONCLUSIONS AND CLINICAL RELEVANCE: The haematology and biochemistry changes observed in cattle infected with T. orientalis Ikeda type were consistent with extravascular haemolytic anaemia. Adult dairy cattle were more likely to be severely anaemic than calves. There were differences in haematology and biochemistry profiles between adult dairy cattle and calves, but most of these differences likely had a physiological rather than pathological basis. Overall, the haematological changes in calves aged <6 months appeared less severe than in adult dairy cattle.     

Increased prevalence of Mycoplasma bovis in the Netherlands

Summary

The epidemiology, therapy, and prevention of M. bovis infections are briefly reviewed In a survey begun in 1982 M. bovis was found frequently in the respiratory of veal calves and beef cattle with respiratory problems. In replacement calves infected with respiratory disease in dairy herds, however, the organism has only been detected since 1986. Respiratory tract specimens collected from calves with respiratory disease were submitted for examination for M. bovis from 1986 to 1991 and originated from 83 herds. Mycoplasma bovis was detected in specimens from 59 of the herds, 20% of which were dairy herds and 80% fattening herds. Arthritis caused by M. bovis was observed in 12 herds until July 1991. Since 1976 when the first mastitis outbreak caused by M. bovis was diagnosed M. bovis has caused 14 more outbreaks. The number of diseased cattle varied from 1 tot 16 per farm, and clinical signs of mastitis varied from mild to severe. In all instances the infection has been eradicated from the herds. Because M. bovis can cause great losses in intensively reared cattle herds, it is advisable to separate purchased veal calves and beef cattle from dairy cattle to prevent further spread of M. bovis.     

The Reproductive Seasons of Some Mammals in the Kruger National Park

During a study on the presence and location of Meloidogyne incognita race 2 within maize roots, a nematode-trapping fungus, Arthrobotrys dactyloides was observed for the first time on maize in South Africa.     

Failure of zinc supplementation to prevent severe facial eczema in cattle fed excess copper

CASE HISTORY: A 400-cow dairy unit in the Waikato suffered a severe outbreak of facial eczema (FE) despite consistent zinc supplementation and significantly elevated serum zinc concentrations.

CLINICAL FINDINGS: FE prevention had begun in mid-January 2006, via zinc sulphate supplementation in the water. Photosensitisation was reported on 06 April, contemporaneous with a marked increase in the number of Pithomyces chartarum spores in pasture samples. Within 10 days of this first clinical case, 100 affected cows had been dried off, eight of which subsequently died or were culled. Blood samples were collected from ten affected cows; all had serum zinc concentrations >17 µmol/L, and eight had gamma-glutamyl transferase (GGT) activities >200 U/ml, indicating that the photosensitisation was secondary to liver damage, i.e. probably FE. Further investigation identified that this herd had been receiving excess copper supplementation; 4/6 culled cows had liver copper concentrations within or above the marginal range for copper toxicity.

DIAGNOSIS: Severe FE despite zinc supplementation that increased serum zinc concentrations above recommended levels

CLINICAL RELEVANCE: As zinc supplementation significantly reduces apparent copper status, copper supplementation is often used to counteract this. Previous excess copper intake may reduce the efficacy of zinc in preventing FE, thus copper intake should be assessed prior to the start of zinc supplementation.     

Isolation and molecular characterisation of Neospora caninum in cattle in New Zealand

AIM: To isolate Neospora caninum from the brains of naturally infected cattle and use molecular techniques to characterise the isolates.

METHODS: Neospora caninum tachyzoites were isolated in Vero cell culture from the brains of a cow and two calves. The isolates were characterised using polymerase chain reaction (PCR) methods, DNA sequencing, an immunofluorescent anti-body test (IFAT), transmission electron microscopy (TEM), and immunohistochemistry (IHC). The brains of the three cattle were subjected to histopathological examination. A pathogenicity study was conducted in 120 BALB/c mice.

RESULTS: Neospora caninum tachyzoites were isolated from all three cases and first observed in vitro between 14 and 17 days post-inoculation. Parasites were sub-cultured and maintained in Vero cell culture for more than 6 months. PCR products were generated for all three isolates, using two different primers. Sequencing of the PCR products and a subsequent BLAST search identified the isolates as N. caninum. In addition, the isolates tested positive using IFAT and IHC, and ultrastructure revealed by TEM was characteristic of N. caninum. Histopathological examination revealed lesions characteristic of N. caninum in 1/3 brains. In the pathogenicity study using BALB/c mice, the mortality rate was 3–7%.

CONCLUSION: This was the first successful isolation of N. caninum in New Zealand confirmed using molecular characterisation tests.     

Rhinotrachéite Infectieuse Bovine Chez Les Vaches Laitières au Québec

An outbreak of infectious rhinotracheitis affecting four herds of dairy cattle was observed in the Province of Quebec, the general characteristics of which are the same as those reported in the infection of feedlot cattle.

However, a 25% average decrease in the milk production of affected herds was noticed during the present infection, thus stressing a particular economic aspect. The nature of the viruses isolated during this outbreak and those described previously in North America is identical.