Fatal cases of Theileria annulata infection in calves in Portugal associated with neoplastic-like lymphoid cell proliferation

This study was carried out to investigate fifteen cases of acute lethal infection of calves (≤ 4 months of age) by the protozoan parasite Theileria (T.) annulata in the south of Portugal. Calves developed multifocal to coalescent nodular skin lesions, similar to multicentric malignant lymphoma. Infestation with ticks (genus Hyalomma) was intense. Theileria was seen in blood and lymph node smears, and T. annulata infection was confirmed by isolation of schizont-transformed cells and sequencing of hypervariable region 4 of the 18S rRNA gene. At necropsy, hemorrhagic nodules or nodules with a hemorrhagic halo were seen, particularly in the skin, subcutaneous tissue, skeletal and cardiac muscles, pharynx, trachea and intestinal serosa. Histologically, nodules were formed by large, round, lymphoblastoid neoplastic-like cells. Immunohistochemistry (IHC) identified these cells as mostly CD3 positive T lymphocytes and MAC387 positive macrophages. A marker for B lymphocytes (CD79αcy) labeled very few cells. T. annulata infected cells in these nodules were also identified by IHC through the use of two monoclonal antibodies (1C7 and 1C12) which are diagnostic for the parasite. It was concluded that the pathological changes observed in the different organs and tissues were caused by proliferation of schizont-infected macrophages, which subsequently stimulate a severe uncontrolled proliferation of uninfected T lymphocytes.     

Status of Lipid Peroxidation,Some Antioxidant Enzymes and Erythrocytic Fragility of Crossbred Cattle Naturally Infected with <Emphasis Type="Italic">Theileria annulata</Emphasis>

Erythrocytic lipid peroxidation, activities of some antioxidant enzymes and osmotic fragility of red blood cells was studied in adult (>1 year) crossbred cattle naturally infected with Theileria annulata. Twenty clinically healthy animals (group I) and 15 clinical cases (group II) of tropical theileriosis were selected. Cattle suffering from theileriosis had significantly higher (p<0.01) erythrocytic lipid peroxidation and osmotic fragility. Activities of antioxidant enzymes, viz. glucose-6-phosphate dehydrogenase (G6PD) and glutamate peroxidase (GPx) were also significantly increased (p<0.01) in group II. However, superoxide dismutase and catalase did not show significant changes. The results indicated that infection with theileria led to increased oxidative stress to the animals, and even a significant rise in the activities of antioxidant enzymes. G6PD and GPx could not lower this oxidative stress. However, the increase in the activities of antioxidant enzymes pointed towards the body’s defence mechanism against lipid peroxidation during oxidative stress in theileriosis.     

牛环形泰勒虫病二温式PCR诊断方法的建立及初步应用

为建立特异、敏感、快速的二温式PCR诊断方法,根据牛环形泰勒虫裂殖子表面抗原(tams1)基因,设计了一对特异性引物,扩增出大小为154bp基因片段,经克隆、测序分析,与已知基因序列的相似性为96%。用建立的牛环形泰勒虫病二温式PCR诊断方法,对从新疆牛环形泰勒虫病流行地区采集的50份全血样品进行诊断,阳性率为88%,而血涂片检出的阳性率只有58%。经试验验证,该方法具有特异性高、敏感性强、重复性和稳定性好等优点。表明本试验所建立的二温式PCR诊断方法可用于牛环形泰勒虫病的临床诊断、隐性感染检测和流行病学调查。     

Seasonal Variation of Boophilus microplus (Canestrini, 1887) (Acari: Ixodidae) in Cattle in Minas Gerais State,Brazil

The seasonal variation of the tick Boophilus microplus in the `Regiaõ Metalúrgica' region of Minas Gerais state was studied using 26 Bos taurus (Holstein)×Bos indicus (Gir) calves, of which 16 animals were 3/4 Holstein×1/4 Gir and 10 were 3/4 Gir×1/4 Holstein. Monthly tick counts were made between March 1996 and March 1998 and, although B. microplus infestations occurred throughout the year, they were greater during the rainy season. Rainfall was the climatic factor that most affected the seasonal variation. The animals with a preponderance of Holstein blood presented infestations that were significantly (p<0.05) greater than those of the other cross. Five of the animals (19.2%) maintained 45.5% of the parasite load of the entire group.     

The Epidemiology of Gastrointestinal Nematodes of Dairy Cattle in Central Kenya

The epidemiology of H. placei and of other gastrointestinal nematodes in yearling dairy cattle was examined on two farms in Kiambu District, central Kenya during each of 13 one-month periods from April 1993 to April 1994. On each farm, 32 newly weaned dairy calves were given a single dose of albendazole and then placed on experimental pastures. Twelve of the animals were designated for bi-monthly slaughter (n = 2) and analysis of worm population characteristics and 20 were designated for blood and faecal collection and for weighing. Two parasite-free tracer calves were grazed alongside the weaner calves each month throughout the study period and were also slaughtered for analysis of worm populations. Faecal egg counts, haematological and serum pepsinogen determinations, herbage larval counts, and animal live weight changes were recorded monthly. The study revealed that Haemonchus placei, Trichostrongylus axei, Cooperia spp. and Oesophagostomum radiatum were responsible for parasitic gastroenteritis and that H. placei was the predominant nematode present in the young cattle on both farms. Faecal egg counts from resident cattle and necropsy worm counts revealed that pasture larval levels were directly related to the amount of rainfall. The total worm burdens in the animals were highest during the rainy season (March–June and October–December) and lowest during the dry seasons (July–September and January–February). The very low recovery of immature larvae of H. placei from the tracer calves indicated that arrested development is not a feature of the life cycle of this parasite in central Kenya. The maintenance of the parasite population depended on continuous cycling of infection between the host and the pasture. The agroclimatic conditions of the study area were such that, in general, favourable weather conditions for the development and survival of the free-living stages of gastrointestinal nematodes existed all year round.     

Impact of Tsetse Control on the Age-specific Prevalence of Trypanosomosis in Village Cattle in Southeast Uganda

The prevalence of trypanosomosis, mean packed cell volume and anti-trypanosomal antibody levels in village cattle of different age groups (<0.5 year, 0.5–2 years, >2–5 years and >5 years) in the areas with tsetse control were compared with those of corresponding age groups in areas without tsetse control in Tororo, southeast Uganda. The prevalence of trypanosomosis in cattle in the age groups of 0.5–2 years, >2–5 years and >5 years in the areas with tsetse control was significantly lower than in cattle in similar age groups in the areas without tsetse control (p<0.5). Trypanosoma vivax was the most predominant Trypanosoma species in the areas with tsetse control, while T. congolense was the most predominant species in the areas without tsetse control. The mean Trypanosoma antibody levels in cattle in the age groups <0.5 year, 0.5–2 years and >2–5 years in the areas with tsetse control were significantly lower than those of the similar age groups in the areas without tsetse control (p<0.5). The mean PCV values for cattle in the age groups 0.5–2 years, >2–5 years and >5 years from the areas with tsetse control were significantly higher than those of the similar age groups in the areas without tsetse control. Tsetse control appeared to have a considerable impact on the prevalence of trypanosomosis, distribution of Trypanosoma species, specific antibody levels and the packed cell volume of cattle in the different age groups.     

Theileria parva: In Vitro Studies on the Effects of Holding Temperature,pH and Medium on Sporozoite Infectivity

The effects of holding temperature, pH and medium on the infectivity of Theileria parva sporozoites were investigated using an in vitro infectivity assay. The sporozoite infectivity lasted for 72 h at a holding temperature of 4 C but for only 24 h at 24 degrees C. Sporozoite infectivity was found to be sensitive to pH variations and sporozoites were most infective between pH 7 and pH 8. There was a significant loss in infectivity at pH 5 and infectivity was almost totally abolished at pH 9. Theileria parva sporozoites are usually held and manipulated in Eagle's minimum essential medium (MEM) with Earles' salts. In this study. Leibovitz-15 supplemented with 15% fetal bovine serum gave a significantly better infectivity than Eagle's MEM (3.8 log units versus 1.0 log units) or any other medium. The importance of proper management of the T. parva sporozoite environment in the laboratory or field is emphasized by the findings in these studies and might also explain some of the failures of vaccination when the pH of the holding medium was allowed to deteriorate.     

Seroprevalence of and Risk Factors for Leptospiral Antibodies among Cattle in the State of Yucatan,Mexico

Sera obtained from cattle in the state of Yucatan, Mexico, were screened using the microscopic agglutination test against 13 serovars of Leptospira interrogans. A total of 62.8% (461/734) cows were positive for one or more serovars. This seroprevalence probably reflects infection because vaccination against leptospirosis has not been practised in Yucatan. The most common antibodies detected were those against antigens of serovars hardjo (54.1%) and tarassovi (53.3%). Region was the only risk factor associated with the seroprevalence of leptospirosis (p < 0.05). This revised version was published online in July 2006 with corrections to the Cover Date.     

Reduction of <Emphasis Type="Italic">Theileria annulata</Emphasis> Infection in Ticks Fed on Calves Immunized with Purified Larval Antigens of <Emphasis Type="Italic">Hyalomma anatolicum anatolicum</Emphasis>

Larval antigen of Hyalomma anatolicum anatolicum, the vector of Theileria annulata, was purified by two-step affinity chromatography using anti-tick gut-specific rabbit IgG and IgG from immunized cattle. The purified antigen showed the presence of a single polypeptide of 37 kDa (GHLAgP) on SDS-PAGE. Two groups (I and II) of naive crossbred calves (Bos taurus × B. indicus) were immunized with 1 mg of GHLAgP in three divided doses. Immunized calves of group I were also infected with a sublethal dose of T. annulata along with a group of non-immunized calves (group III). Animals in groups I, II, III as well a control group (group IV) were challenged with live nymphs of H. a. anatolicum on the 10th day of immunization. There was a significant reduction in the number of emerging adults of 56.9% ± 1.67% in calves of group I (p < 0.01) and 63.09% ± 1.26% in calves of group II (p < 0.001) compared to the controls. The calves of groups I and II showed antibody responses to tick antigen up to day 70 post immunization. Infection with T. annulata was determined in the salivary glands of adult ticks that developed from the nymphs used for challenge infection. In ticks taken from group I calves, there was a 75.0% ± 0.00% infection compared with only 85.0% ± 2.88% infection in ticks taken from calves of group III. Using PCR, a lower infection (83.33% ± 3.33%) was detected in ticks that developed from calves of group I compared with calves from group III (90.00% ± 2.88%). The ground-up tick supernatants (GUTS) of the ticks taken from calves of group III yielded higher infection rate and exhibited higher infectivity titre in in vitro infection assay of bovine mononuclear cells than the GUTS of the ticks taken from calves of group I. The results suggest a partial reduction in growth rate of T. annulata in ticks feeding on calves immunized with GHLAgP.     

Lechiguana (Focal Proliferative Fibrogranulomatous Panniculitis) in Cattle

Lechiguana is a disease of cattle characterized by large, hard, subcutaneous swellings that grow rapidly and result in death after 3–11 months in untreated animals. Cattle treated with antibiotics recover. The disease has been reported from five states in south and southeastern Brazil. Histologically, the lesion consists of focal proliferation of fibrous tissue infiltrated by plasma cells, eosinophils, lymphocytes and sometimes neutrophils. The primary lesion is an eosinophilic lymphangitis, which results in eosinophilic abscesses, with occasional rosettes containing bacteria in their centres. Much experimental and epidemiological evidence, reviewed in this article, supports the suggestion that lechiguana is caused by an association of Pasteurella granulomatis (syn: Mannheimia granulomatis) and Dermatobia hominis.     

Clostridium Infection (Jisizheng) in Yaks in Qinghai,China

Since the mid-1980s, outbreaks of a disease characterized by a sudden onset, acute deaths and extensive haemorrhages in the viscera and digestive tract of yaks have been prevalent in Qilian, Qinghai, China. The disease is known as jisizheng by local people. Virulent Clostridium perfringens type A and Clostridium haemolytica were isolated from yaks that had died of jisizheng. In 1996 and 1997, yaks were immunized with a polyvalent inactivated vaccine against C. perfringens and with an inactivated vaccine against C. haemolyticum, and this prevented the occurrence of jisizheng.     

The Prevalence and Intensity of Helminth and Coccidial Infections in Dairy Cattle in Central Kenya

A survey of gastrointestinal parasite infections of young (<6 months old), immature (6–12 months old) and adult (>12 months old) dairy cattle on 16 farms in Kiambu District, Kenya was conducted during a dry season (September 1991 to January 1992) and during a wet season (March to July 1992). The survey was based on monthly coproparasitological examination of cohorts and worm counts in tracer calves. The effects of age, sex, farm and season on the prevalence and intensity of helminth and coccidial infections were determined. Faecal egg and oocyst counts revealed that the overall prevalences were: strongyles (including trichostrongyles) (85.5%), liver flukes (Fasciola gigantica) (34.0%), coccidia (30.9%) and tapeworms (9.6%). Eight species of the protozoan Eimeria were identified, the most prevalent species being E. bovis and E. zuernii. The most prevalent nematode genera were Haemonchus, Cooperia, Oesophagostomum and Trichostrongylus. Season, farm and age of the animals had a significant (p<0.05) influence on the intensity of infection with strongyles, liver flukes and coccidia, whereas the sex of the animals had no significant (p>0.05) effect on the prevalence or intensity of infections. A higher intensity of infection with strongyles and coccidia was found in the wet season than in the dry season (p<0.05). The age-specific intensity was in the following order: for strongyles, immature animals of 6–12 months of age had the highest egg counts, followed by young calves and adults. Calves had significantly (p<0.05) higher oocyst counts than immatures or adults. Liver fluke egg counts did not differ significant (p>0.05) between immatures and adult cattle.     

Epidemiology of the epidemic of bovine anaemia associated with Theileria orientalis (Ikeda) between August 2012 and March 2014

AIMS: To describe the epidemiology of the epidemic of bovine anaemia associated with Theileria orientalis infection (TABA) in New Zealand between 30 August 2012 and 4 March 2014.

METHODS: Blood samples and associated data were obtained from cases of TABA. The case definition for TABA was met when piroplasms were present on blood smears and the haematocrit was ≤0.24?L/L. Samples were analysed using quantitative PCR (qPCR) assays for the detection of T. orientalis Ikeda type. Only cases that were positive in the qPCR assays were included in the analysis. A case herd was defined as a herd that had ≥1 animal positive for T. orientalis Ikeda.

Movement records for farms were accessed through the national animal identification and tracing scheme. The OR for cattle movements onto a case farm compared to a non-case farm was estimated using a generalised estimating equation model and the geodesic distance for movements onto case and non-case farms compared using Student's t-test. The kernel-smoothed risk of disease at the farm level was calculated using an extraction map and the clustering of diseased farms in time and space was measured using the spatial temporal inhomogeneous pair correlation function.

RESULTS: In the first 18 months there were 496 case herds; 392 (79%) were dairy and 104 (21%) beef herds. Of 882 individual cases, 820 (93.0%) were positive for T. orientalis Ikeda in the qPCR assays. Case herds were initially clustered in the Northland, then the Waikato regions. The OR for a case farm compared to a non-case farm having ≥1 inward cattle movements was 2.03 (95% CI=1.52–2.71) and the distance moved was 26 (95% CI=20.8–31.3) km greater for case farms. The risk of disease was highest in a north, north-eastern to south, south-western belt across the Waikato region. The spatial-temporal analysis showed significant clustering of infected herds within 20–30 days and up to 15?km distant from a case farm.

CONCLUSIONS: Theileria orientalis Ikeda type is likely to have been introduced into regions populated with naïve cattle by the movement of parasitaemic cattle from affected areas. Local spread through dispersed ticks then probably became more important for disease transmission between herds once the disease established in a new area.

CLINICAL RELEVANCE: Dairy and beef farming in the North Island of New Zealand will be significantly changed in the coming years by the incursion of this new disease.     

Investigations on the Prevalence of Bovine Tuberculosis and Brucellosis in Dairy Cattle in Dar es Salaam Region and in Zebu Cattle in Lugoba Area,Tanzania

A study between August 1995 and December 1997 included 343 dairy cattle on 20 farms in the Dar es Salaam region and 2289 zebu cattle on 39 bomas in the Lugoba area (coast region). The aim was to establish the prevalence of bovine tuberculosis (Mycobacterium bovis) and bovine brucellosis (Brucella abortus). In the single intradermal tuberculin test (SIT), 0.9% (3/343) of the animals in Dar es Salaam tested positive and 1.2% (4/343) were doubtful. Positive reactors were found in 10% (2/20) of the farms. In the Lugoba area, 0.6% (14/2206) were positive and 6.8% (149/2206) doubtful, positive cases being found in 21% (8/39) of all bomas. In the slow agglutination test (SAT) for B. abortus, 14.1% (48/341) of the serum samples reacted positively in Dar es Salaam and 2.3% (8/341) were doubtful. Positive SAT reactors were identified on 25% (5/20) of the dairy cattle farms. In the Lugoba area, 12.3% (273/2221) proved to be positive SAT reactors and doubtful reactions were observed in 2.9% (64/2221). SAT-positive animals were detected on 87% (34/39) of all bomas. The prevalence in single herds in Dar es Salaam varied from 4.3% to 5.3% for the SIT and from 2.2% to 50% for the SAT. The prevalence in single herds in Lugoba area was between 1.1% and 2.9% for SIT and from 1.4% up to 62.1% for SAT. The two cattle populations differed significantly (p<0.001) in the prevalence of both bovine tuberculosis and bovine brucellosis. Two cows that were positive reactors were slaughtered and subjected to post-mortem examination, and organ samples were bacteriologically cultured. The occurrence of Mycobacterium tuberculosis was confirmed by polymerase chain reaction (PCR) in both cows.     

杂交改良对云南地方黄牛屠宰性能的影响研究

本试验研究了全放牧饲养条件下云南地方黄牛和西门塔尔与云南地方黄牛杂交F1代的屠宰性能。试验屠宰6头地方黄牛和8头杂交牛。结果显示,杂交F1代与地方黄牛相比,体高、体斜长、十字部高、尻长、坐骨端宽、胸宽、管围、后腿围和腹围显著提高(P<0.05);宰前活重、胴体重、净肉重、骨重和生殖器周围脂肪重显著增加(P<0.01);胴体长(P<0.01)、胴体后腿围(P<0.01)、胴体后腿宽(P<0.01)和胴体深(P<0.05)显著提高,但胴体后腿长显著降低(P<0.01)。本试验结果表明,利用国外优良肉牛品种西门塔尔与云南地方黄牛杂交有效地改善了杂交牛的生产性能和产肉性能。     

Influence of Age and Breed on Natural Bovine Fasciolosis in an Endemic Area (Galicia,NW Spain)

An analysis was undertaken of the effect of breed and age on bovine fasciolosis using antibody and antigen detection ELISAs. A total of 84.3% of the 1284 serum samples examined had positive antibody values and 20.4% exhibited antigenaemia. The seroprevalence of antibodies in crossbred cattle was higher than that in autochthonous Rubia Gallega, Friesian or Brown Swiss cows. The highest percentage antigenaemia occurred in the Brown Swiss cattle, but the breed differences were not statistically significant. Cattle aged 3–5 years had the highest antibody and circulating antigen prevalence and the age differences were significant. It was concluded that the apparent influence of breed was probably closely associated with the husbandry system. The autochthonous Rubia Gallega may be better adapted to fasciolosis as its percentage of antigenaemia was the lowest.     

Comparison of the Standard AGID Test and Competitive ELISA for Detecting Bluetongue Virus Antibodies in Camels in Gujarat,India

The performance of the standard agar gel immunodiffusion (AGID) test and the competitive enzyme-linked immunosorbent assay (cELISA) for the detection of serum antibody against bluetongue virus (BTV) in clinically healthy and diseased camels in Gujarat state were compared. Out of 176 sera tested, 22 (12.5%) and 34 (19.3%) were positive for group-specific bluetongue antibodies by AGID and cELISA, respectively. Maximum seropositivities of 18.0% by AGID and 25.8% by cELISA were recorded in the Kutchhi breed, and of 6.9% and 12.6%, respectively, in the Marwari breed. The seroprevalence detected by AGID and cELISA in clinically healthy and diseased camels did not differ significantly with regard to bluetongue disease in these breeds.     

Treatment of East Coast Fever of Cattle with a Combination of Parvaquone and Frusemide

Pulmonary oedema is a common sign of East Coast fever (ECF, Theileria parva infection) of cattle. A trial was conducted on farms in Uganda to compare a product containing both the antitheilerial compound parvaquone and the diuretic compound frusemide with one containing only parvaquone, in the treatment of ECF. The trial involved 40 clinical cases of ECF, some of them complicated by other infections, in cattle of all ages and on several farms. Confirmed cases were treated with either parvaquone+frusemide (P+F) or parvaquone alone (P). Survival after treatment with P+F was 77% compared with 71% with P. Five of the 10 fatalities were complicated cases. The cure rate for severe but uncomplicated ECF was 89% with P+F and 40% with P. Pulmonary signs were resolved within 24-48 h after treatment with P+F and clinical recovery was noticeably more rapid than with P. The antiparasitic effect of the two treatments was similar. P+F could be particularly useful when reporting, diagnosis or laboratory confirmation of ECF is delayed, because advanced cases are more likely to be encountered under these circumstances.