Anemia In Cattle Research Articles

Determining diagnostic sensitivity loss limits for sample pooling in duplex rtPCR surveillance testing: Theileria orientalis and Anaplasma marginale.

To expand surveillance testing capacity through sample pooling, a thorough understanding is needed of how sample dilution through pooling affects the sensitivity of candidate assays. We validated a robust and representative framework for assessing the dilution effect of sample pooling using duplex rtPCR surveillance of Theileria orientalis and Anaplasma marginale, both of which are causative agents of severe anemia in cattle and a serious threat to the cattle industry in Virginia and many other states. We used 200 known-positive samples with Ct values representative of typical surveillance results in a series of pools in which we re-tested each sample individually, followed by each sample diluted in equal volumes with negative samples to make pools of 2, 4, 6, 8, and 10 total samples. We compared the Ct values of the individual positives with the Ct values of each pool size to determine if Ct values increase past the limit of detection in the 45-cycle assay. We observed a maximum of 2% sensitivity loss (no more than 2 of 100 samples returned a false-negative result) for both T. orientalis and A. marginale during the pooling series, with lower-than-expected average Ct increase and sensitivity loss. We conclude that pooling up to 10 samples would be acceptable for regional surveillance of T. orientalis and A. marginale using our rtPCR assay. The described strategy is applicable to validate pooling for a wide range of single and duplex rtPCR assays, which could expand efficient disease surveillance.

Development of a Specific PCR Assay for Theileria sp. Yokoyama and Assessment of Its Potential to Cause Anemia in Cattle.

The clinical implications of Theileria sp. Yokoyama, a recently identified Theileria species in cattle, remain uncertain. The objective of the present study was to evaluate the anemia status in cattle infected with Theileria sp. Yokoyama. Blood samples were collected from 206 cattle across seven Veterinary Ranges in Sri Lanka and analyzed for red blood cell (RBC) indices, including hemoglobin concentration, hematocrit, and RBC counts. Additionally, DNA was extracted from the samples and screened with a newly developed Theileria sp. Yokoyama-specific PCR assay targeting the cytochrome b gene. The PCR results revealed that 60 (29.1%) of the surveyed cattle tested positive for Theileria sp. Yokoyama, with 47 (78.3%) of them being co-infected with other hemopathogen species. Our findings revealed that the cattle breeds, management systems, and tick infestations are potential risk factors for the Theileria sp. Yokoyama infection. Next, we evaluated the anemia status among the surveyed cattle based on the RBC indices. We found that all non-infected cattle were non-anemic. By contrast, anemia was observed in 15 Theileria sp. Yokoyama-infected cattle, including 3 singly infected (anemia rate 3/13, 23.1%) and 12 co-infected cattle (12/47, 25.5%). Our findings suggest that Theileria sp. Yokoyama causes anemia in infected cattle.

Evaluation of haematological parameters in haemolytic anaemia caused by tick-borne pathogens in grazing cattle.

No tick-borne pathogens (TBPs) causing haemolytic anaemia in cattle have been reported, except Theileria orientalis and complete blood count (CBC) profile is the only haematological parameter to determine the severity of regenerative haemolytic anaemia. To identify the causative agents of TBP-induced haemolytic anaemia and determine haematological parameters that indicate haemolytic anaemia in grazing cattle. Eighty-two Korean indigenous cattle (Hanwoo) were divided into two groups: grazing (n = 67) and indoor (n = 15) groups. CBC and serum biochemistry were performed. PCR was conducted using whole blood-extracted DNA to investigate the prevalence of TBPs. TBP-induced haemolytic anaemia was observed in the grazing group. In grazing cattle, co-infection (43.3%, 29/67) was most frequently detected, followed by T. orientalis (37.6%, 25/67) and Anaplasma phagocytophilum infections (1.5%, 1/67). In indoor cattle, only co-infection (20%, 3/15) was identified. Grazing cattle exhibited regenerative haemolytic anaemia with marked monocytosis, mild neutropenia, and thrombocytopenia. According to grazing frequency, the 1st-time grazing group had more severe anaemia than the 2nd-time grazing group. Elevations in indirect bilirubin and L-lactate due to haemolytic anaemia were identified, and correlations with the respective markers were determined in co-infected grazing cattle. Quantitative evaluation of haematocrit, mean corpuscular volume, and reticulocytes (markers of regenerative haemolytic anaemia in cattle) was performed for the first time. Our results show that, in addition to T. orientalis, A. phagocytophilum is strongly associated with anaemia. The correlation between haemolytic anaemia severity and haematological parameters (indirect bilirubin, reticulocytes, and L-lactate) was confirmed.

Examination of Bovine Red Blood Cell Death in Vitro in Response to Pathophysiologic Proapoptotic Stimuli.

Interspecies variations in mammalian red blood cells (RBCs) are observed in circulating RBC lifespan, cell size, fluidity, aggregation, water permeability, metabolism, lipid composition, and the overall proteome. Bovine RBC cell membrane is deficient in phosphatidylcholine and exhibits anomalies in the arrangement of phosphatidylethanolamine within the lipid bilayer. However, like human RBCs, virtually all the aminophospholipid phosphatidylserine (PS) is found within the cytoplasmic side of the cell membrane of intact circulating bovine RBCs. During apoptotic cell death of human and murine RBCs, PS translocates to the outer leaflet of the cell membrane via Ca2+-dependent and -independent signaling mechanisms. However, little is known about this process in bovine RBCs. Using cytofluorometry analyses, we characterized and compared the cell death responses in bovine and human RBCs in vitro exposed to various pathophysiologic cell stressors. Ionic stress, by ionophore treatment, and oxidative stress enhanced cytoplasmic Ca2+ levels and cell membrane PS expression in both bovine and human RBCs. Fever-grade hyperthermia and energy starvation promoted Ca2+ influx and elevated reactive oxygen species levels in both human and bovine RBCs. However, bovine RBCs displayed minimal increases in PS expression elicited by hyperthermia, energy starvation, and extracellular hypertonicity as compared to human RBCs. In response to decreased extracellular osmolality, bovine RBCs exhibited significantly enhanced fragility as compared to human RBCs. Bovine RBCs display differential cell death patterns as compared to human RBCs, only partly explained by increased Ca2+ influx and oxidative stress. Premature removal of circulating RBCs could potentially contribute to the pathogenesis of anemia in cattle caused by a wide range of factors such as systemic diseases, parasitic infections, and nutritional deficiencies.

Trypanosome Infections and Anemia in Cattle Returning from Transhumance in Tsetse-Infested Areas of Cameroon.

The objective of this work was to assess the anemic status and the use of an immunological test and PCR-based methods to determine the infection rates of trypanosomes species. Transhumance aims to provide cattle with greener pastures and greater water resources than in the Djerem region during the dry season. Two criteria were used to assess the health status of the animals, the prevalence of trypanosomiasis and the level of anemia. In addition, we have evaluated the effectiveness, in trypanosomiasis detection, of the Very Diag Kit (CEVA Santé animale), a Rapid diagnosis test (RDT) based on immunological identification of T. congolense s.l. and T. vivax, responsible for AAT. Four trypanosome species (Trypanosoma congolense savannah type (Tcs), T. congolense forest type (Tcf), T. brucei s.l. (Tbr) and T. vivax (Tvx)) were identified in cattle sampled in four villages. The overall infection rate determined by PCR (68.6%) was much higher than those generally reported in cattle from the Adamawa region (35 to 50%). Infections (including mixed infections) by Tc s.l. (Tcs + Tcf) were predominant (45.7%). The infection rates were also determined using the Very Diag Kit allowing us to identify Tc s.l. and Tvx in the field in less than 20 min. This method provided, for the global infection, a higher rate (76.5%) than that determined by PCR (68.6%), although it is supposed to be less sensitive than PCR. Tc s.l. infection rate (37.8%) was similar to that (38.8%) determined by PCR (Tcs + Tcf single infections). In contrast, the prevalence of Tvx single infections measured by RDT (18%) was nearly two-fold higher than that (9.4%) measured by PCR. Thus, further comparative analyses seem to be needed in order to more accurately assess the sensitivity and specificity of the Very Diag test under our conditions of use on blood samples. The mean PCVs in trypanosome-infected as well as in uninfected cattle were below 25%, the threshold below which an animal is considered anemic. Our study shows that cattle return from transhumance in poor health. It raises questions about its real benefit, especially since the herds are themselves likely to become vectors of trypanosomiasis and possibly of other diseases. At least, effective measures have to be undertaken to treat all cattle coming back from transhumance.

Efficacy of Whole Blood Transfusion as an Adjunct Therapy in Severe Cases of Haemolytic Anaemia in Cattle

Background: In view of common occurrence of haemoprotozoan infections leading to severe haemolytic anaemia and increased case fatality rate in cattle in study area the present investigation was planned to evaluate efficacy of whole blood transfusion as an adjunct therapy in severe cases of hemolytic anaemia in cattle. Method: Sixteen cattle suffering from one or other haemoprotozoan infections induced haemolytic anaemia and having haemoglobin values less than 5 gm/dl were randomly divided in to two treatment groups. Group A (n=8) cattle were treated with standard treatment of haemoprotozoan infection without blood transfusion while group B (n=8) cattle were treated with whole blood transfusion in addition to standard treatment for haemoprotozoan infections. Result: Clinical and haematological parameters revealed rapid improvement in group B cattle as compared to group A cattle. The survival rate was significantly higher in group B cattle (100%) as compared to group A cattle (62.5%). The haemoglobin and haematocrit values were also found significantly higher in group B as compared to group A at 12 hours as well as on 21st day post treatment. Thus, it is concluded that, whole blood transfusion could be used as an adjunct therapy to hasten clinical recovery and to increase the survivability in cattle suffering from severe haemolytic anaemia due to haemoprotozoan infections.

Prevalence of Bovine Trypanosomosis and Its Associated Risk Factor in Hawa Galan District, Kelem Wollega Zone of Ethiopia.

An analytical cross-sectional study was performed between November 2015 and April 2016 at Hawa Galan woreda, Kelem Wollega Zone, Oromia Region, Ethiopia, to estimate the proportion of cattle with trypanosomosis and to evaluate the elements associated with the prevalence of bovine trypanosomosis. A haemoparasitological examination of the Buffy coat method was implemented to determine the proportion of trypanosomosis-positive cattle. A study population of 428 bovines was randomly selected from three peasant associations (PAs) and tested for the presence of the disease. Findings of the laboratory results indicate that among all animals tested at the study time, 26 (6%) animals were positive for the disease. Among the positive results, the proportion of trypanosome species was Trypanosoma congolense 18 (69.3%) and Trypanosoma vivax 8 (30.7%). The prevalence at the selected PAs was 12.5%, 3.8%, and 3% for Lemlem, Madawalkituma, and Ifajiru, respectively. From this finding, the relationship has a statistically significant variation (P < 0.05) among peasant associations and body condition state, and the proportion was significantly greater (P < 0.05) in animals categorized under poor body condition score. The relationships between age and sex of cattle show no statistically significant variation among them (P > 0.05). This study indicates that the proportion of trypanosomosis was greater in anemic (PCV<24%) cattle than nonanemic (PCV≥24%). Therefore, based on this finding, the proportion of bovine trypanosomosis is evident that can cause a major effect on the health of cattle in the study population in the study area. So, effective control methods could be applied to decrease the disease and its related economic loss.

Theileria orientalis outbreak in an organized cattle breeding farm.

Theileriosis is an important tick borne disease of cattle caused by a haemoprotozoan of genus Theileria. Clinical bovine theileriosis is mainly caused by T. annulata or T. parva but the clinical disease due to T. orientalis is rare. T. orientalis mainly infect RBCs and causes "Oriental theileriosis" or Theileria-associated bovine anaemia in cattle and other livestock species. Two genotypes of T. orientalis (Chitose and Ikeda) are reported to cause severe disease in some countries. In this report, a spontaneous outbreak of Oriental theileriosis was studied in an organized Holstein-Friesian cattle breeding farm situated in the south-eastern Himalayan ranges of Himachal Pradesh State of India. Animal blood and tick samples were tested using cytological and PCR techniques. The disease episode occurred in a protracted manner spanning over 10 to 12 months and association of T. orientalis was confirmed in 93.3% of the blood and 21.7% of Rhipicephalus microplus (tick) samples. No other tick borne pathogen was detected except Anaplasma marginale in two blood samples. Haematological profiling of infected cattle showed characteristic indicators of anaemia like haemoblobin, RBC count, haematocrit value and mean corpuscular volume at either lower than normal or near the lower normal range. The prevailing persistent anaemic changes led to more severe clinical manifestations like abortion and joint inflammation. The detected T. orientalis strains and ticks species were further confirmed by nucleotide sequence analysis of 18S rRNA and 16S rRNA genes. Phylogenetically, T. orientalis strains showed clustering with other reported strains of T. orientalis from the surrounding regions. This first report of clinical Oriental theileriosis from India emphasises the importance of T. orientalis as an emerging tick borne pathogen and role of widely prevalent ticks species in disease transmission and their impact on livestock production.

Molecular identification of trypanosomes in cattle in Malawi using PCR methods and nanopore sequencing: epidemiological implications for the control of human and animal trypanosomiases.

This study aimed to identify trypanosomes infecting cattle in Malawi in order to understand the importance of cattle in the transmission dynamics of Human African Trypanosomiasis (HAT) and Animal African Trypanosomosis (AAT). A total of 446 DNA samples from cattle blood from three regions of Malawi were screened for African trypanosomes by ITS1 PCR. The obtained amplicons were sequenced using a portable next-generation sequencer, MinION, for validation. Comparison of the results from ITS1 PCR and MinION sequencing showed that combining the two methods provided more accurate species identification than ITS1 PCR alone. Further PCR screening targeting the serum resistance-associated (SRA) gene was conducted to detect Trypanosoma brucei rhodesiense. Trypanosoma congolense was the most prevalent Trypanosoma sp., which was found in Nkhotakota (10.8%; 20 of 185), followed by Kasungu (2.5%; 5 of 199). Of note, the prevalence of T. b. rhodesiense detected by SRA PCR was high in Kasungu and Nkhotakota showing 9.5% (19 of 199) and 2.7% (5 of 185), respectively. We report the presence of animal African trypanosomes and T. b. rhodesiense from cattle at the human–livestock–wildlife interface for the first time in Malawi. Our results confirmed that animal trypanosomes are important causes of anemia in cattle and that cattle are potential reservoirs for human African trypanosomiasis in Malawi.

A Study on Prevalence of Trypanosomosis, its Risk Factors and Anaemia in Cattle of Damot Woyde District, Southern Ethiopia

A Study on Prevalence of Trypanosomosis, its Risk Factors and Anaemia in Cattle of Damot Woyde District, Southern Ethiopia

Basophilic stippling and reactive lymphocytosis in bovine anaplasmosis - A case report

Anaplasmosis is known for causing anemia in cattle. However, morphological changes in the red and white blood cells are not often described in bovine anaplasmosis cases. Thus, the aim of this work was to describe changes in blood cell morphology, such as the incidence of basophilic stippling in red blood cells and the reactive lymphocytosis, presented by a 10-month-old Jersey heifer, who was bred in a dairy farm in Juiz de Fora County (MG), Forest Zone of Minas Gerais State (Zona da Mata Mineira). The animal presented clinical signs of hyperthermia and hypocorous mucosae, which suggested hemoparasitosis. Laboratory examination demonstrated the presence of Anaplasma marginale in several erythrocytes, 13.4% of parasitemia, red blood cells presenting basophilic stippling, severe macrocytic normochromic anemia, increased total number of leukocytes resulting from the increased number of lymphocytes, some large lymphocytes with basophilic cytoplasm, and other lymphocytes showed granules in the cytoplasm. These findings are not often described in bovine anaplasmosis cases. So, this work shows the relevance of blood smear examinations to diagnose the hemoparasite, as well as to identify cellular morphological changes caused by it.

Prevalence studies for Theileria orientalis conducted during the early stages of the 2012 New Zealand epidemic of Theileria associated bovine anaemia.

Since 1982 there have been two epidemics of bovine anaemia associated with Theileria orientalis infection (TABA) in New Zealand. The latest and more serious epidemic started in 2012 and is associated with the T. orientalis (Ikeda) type. In 2013 the Ministry for Primary Industries (MPI) carried out several epidemiological investigations on the TABA epidemic, which included a series of on-farm herd prevalence studies. The aims of this paper are to present and summarise the results of these herd prevalence studies. The data were collected from March 2013 to September 2013 and comprised 26 farms. For 21/26 of these farms, a T. orientalis (Ikeda) positive index case of TABA precipitated the follow-up herd prevalence study. Each herd prevalence study involved a random sample of 10 cows for haematocrit, for piroplasm counts (number of Theileria infected RBCs per 1000 RBCs) and for PCR molecular diagnosis. Animals were diagnosed anaemic if the haematocrit ≤0.24. The results showed that for the index cases, the average haematocrit = 0.10 L/L and the average piroplasm count = 10.9 Theileria infected RBCs per 1000 RBCs, and for the herd prevalence studies the average haematocrit = 0.26 and the average piroplasm count = 6.9. The average herd prevalence of T. orientalis (Ikeda) PCR positive cattle and of anaemic cattle was 87% and 26% respectively. For dairy herds (n = 7 herds) it took, on average, 80 days for the within herd prevalence of anaemia to return to zero after the initial diagnosis. In conclusion the prevalence studies showed that infection rates and anaemia rates were already high in herds when the index case of TABA was observed. At the time, these prevalence studies significantly added to our understanding of the epidemiology of the evolving T. orientalis (Ikeda) epidemic and furthermore met the New Zealand MPI objective of being rapid and applied, fit for purpose and completed at a relatively low cost.

Clinical, hemato-biochemical alterations and oxidant–antioxidant biomarkers in Babesia-infected calves

Babesia is one of the main causes of anemia in cattle, a lot of elucidations have been suggested to explain its pathogenesis. This study was designed to investigate clinical, hemato-biochemical and oxidant/antioxidant status and its relation with the resultant anemia in Babesia-infected calves. Seventeen (17) native breed calves were involved in this study, clinical signs and microscopic findings were recorded, also blood samples were taken to investigate hematologic changes, serum biochemical variations and oxidative stress biomarkers. The most commonly observed clinical signs were fever, emaciation, depression, icterus and hemoglobinuria. Significant reduction in PCV, HB, RBCs, MCHC, Total protein, and albumin along with significant increase in MCV, WBCs, monocytes and BUN were the most consistent hemato-biochemical changes. Oxidant/antioxidant and trace mineral assessment showed significant reduction in Superoxide dismutase “SOD”, Glutathione peroxidase “GPx”, Zn, Cu along with significant increase in malondialdehyde (MDA) activities. In the current investigation, oxidant/antioxidant imbalance along with the synchronized alterations in antioxidant trace minerals was detected in Babesia-infected calves. These findings support notion that Babesia infection associated with oxidative stress and this process may be linked to the resultant anemia.

Prevalence of pathogenic trypanosomes in anaemic cattle from trypanosomosis challenged areas of Itezhi-tezhi district in central Zambia.

BackgroundThe measure of anaemia status using packed cell volume (PCV) can be a reliable indicator of African trypanosomosis (AT) in the absence of other anaemia-causing conditions. However, studies that have estimated prevalence of anaemia in cattle from AT endemic areas have rarely reported the prevalence of the disease in the anaemic cattle. Therefore we investigated the prevalence of AT in anaemic cattle at sites that had recently reported the disease in Itezhi tezhi district of central Zambia.MethodsDuring a survey, blood samples were collected from 564 randomly selected cattle for anaemia determination from seven crush pens (Mutenda, Kapulwe, Banachoongo, Itumbi, Iyanda, New Ngoma and Shinampamba). At a PCV- value cut off of 26 %, all samples positive for anaemia were subjected to both parasitological examination on thick and thin blood smears and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) for detection of trypanosome DNA. Fisher’s exact test and a mixed effect logistic regression analyses were used to determine and measures associations, respectively.ResultsOf 564 cattle screened, 58 (10.3 %; 95 % CI: 7.8–12.8 %) had anaemia. PCR-RFLP results showed that 17 (29.3 %; 95 % CI; 17.2–41.4 %) anaemic cattle were positive for pathogenic trypanosomes compared to 1 (1.7 %; 95 % CI: 0.0–5.2 %) on parasitological examination using thick smears. The infections were caused by Trypanosoma congolense and Trypanosoma vivax. Fisher’s exact test showed a strong association between PCV and pathogenic trypanosome infection (P = 0.004). A mixed effect multivariate logistic regression showed that a one unit increase in PCV reduced the likelihood of detecting AT with PCR-RFLP by 24.7 % (95 % CI: 4.6–40.6 %; P = 0.019) in anaemic cattle, taking into account their age and sex, with random effects for crush pen.ConclusionThese results suggest that T. congolense and T. vivax could be important causes of anaemia in cattle reared in AT endemic areas of Itezhi tezhi in Central Zambia. This also suggests that even though pathogenic trypanosomal infection was strongly associated with PCV, it could only account for up to 41 % of the anaemia in cattle. Therefore further investigation to ascertain other factors responsible for anaemia in AT endemic areas of Itezhi tezhi in Central Zambia is needed.

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.

Application of quantitative PCR assays for diagnosing Ikeda and other Theileria orientalis types to examine associations between severity of anaemia and parasitaemia in bovine anaemia outbreaks

AIMS: To use quantitative PCR assays to detect Theileria orientalis Ikeda type in cattle presumed infected with T. orientalis, to examine the relationship between theilerial piroplasm count and haematocrit (HCT), and the relationship with quantification cycle threshold (Cq) values.METHODS: Blood samples in EDTA (n=1,024), derived from herds affected by anaemia associated with T. orientalis infection (TABA) between April and October 2013, were submitted for testing using quantitative PCR (qPCR) assays for T. orientalis and Ikeda type. Nucleotide sequencing of the major piroplasm surface protein (MPSP) gene was performed on 16 samples to identify T. orientalis types. Blood smear and/or HCT results were supplied with most samples. For data analysis, the number of theilerial piroplasm per 1,000 erythrocytes counted was categorised as negative (0), low (1–9), moderate (10–100) or high (>100). HCT was categorised as severely anaemic (<0.15 L/L), mildly anaemic (0.15–0.24 L/L) or not anaemic (>0.24 L/L). Differences between categories in proportion of samples positive for Ikeda type or mean Cq value were examined using χ2 tests or analysis of variance, respectively.RESULTS: Of 1,022 samples containing amplifiable DNA, 916 (90%) were positive for T. orientalis and 789 (77%) were positive for Ikeda type. Nucleotide sequencing of MPSP amplicons also identified the presence of Chitose and Buffeli types in 11 samples without Ikeda. Ikeda was detected in a greater proportion of severely anaemic (288/302; 95%) than mildly anaemic (227/252; 90%) cattle (p=0.02). In non-anaemic cattle, 344/406 (85%) were positive for T. orientalis and 247/406 (60%) were positive for Ikeda type. In samples from cattle that were piroplasm-positive, a greater proportion of anaemic (483/505, 96%) than non-anaemic (211/307; 69%) cattle were positive for Ikeda type (p<0.001). In piroplasm-negative cattle, 20/37 (54%) anaemic and 25/78 (32%) non-anaemic cattle were Ikeda-positive (p<0.05). The distributions of Cq values differed between piroplasm count and HCT categories (p<0.001). Mean Cq differed between high and negative, and low piroplasm categories (p<0.001), but not between high and moderate categories (p=0.81), and differed between severely anaemic and mildly anaemic (p<0.001), and non-anaemic categories (p<0.001).CONCLUSIONS: The Ikeda type was found in a high proportion of cattle during outbreaks of TABA in New Zealand. Analysis of Cq values suggested a relationship of Ikeda parasitaemia with severity of anaemia, but further investigation is required to better understand the role of parasitaemia in the pathogenesis of TABA.

Effeciency of Various Supportive Treatments as A Cure for Anaemia in Cattle with Theileriosis

Effeciency of Various Supportive Treatments as A Cure for Anaemia in Cattle with Theileriosis

Etiology and haemato-biochemical alterations in cattle of Jammu suffering from anaemia

Aim: To obtain the clinical appraisal and haemato-biochemical alterations in cattle suffering from anaemia in Jammu division of J&K, India. Materials and Methods: 125 cattle were screened for anaemia on the basis of Haemoglobin (Hb) concentration. Blood samples were taken for estimation of Hb, packed cell volume (PCV), total erythrocyte count (TEC), mean corpuscular volume (MCV), mean corpuscular haemoglobin (MCH), mean corpuscular haemoglobin concentration (MCHC), total protein, albumin, total bilirubin, plasma iron and total iron binding capacity (TIBC). Faecal samples were screened for ova/ cyst and peripheral blood smear was examined for haemoprotozoa. The animals with Hb < 8g/ dl were categorized as anaemic. Results: Out of 125 cattle screened for anaemia, 47 (37.6%) were anaemic. Levels of Hb, PCV, TEC, MCH, MCHC and plasma iron were significantly lower in anaemic cattle than non-anaemic cattle while TIBC was significantly higher. The clinical signs in majority of the anaemic animals were depression, pale mucous membrane, dehydration and rough body coat. Faecal examination revealed parasitic infestation in 25 of those anaemic animals (Trichostrongyloids - 16 and Bunostomum09). Ticks were seen in seven animals, whereas three animals were lice/ flea infested. Peripheral blood smear examination revealed Babesia bigemina infection in five animals. Conclusion: Hb, PCV, Plasma iron and TIBC were significantly altered in the present study and can be used for diagnosis of anaemia in cattle. The major causes of anaemia in this study were endo- and ecto-parasitic infestations.

Isolation, cloning, and pathologic analysis of Trypanosoma evansi field isolates

In recent years, the emergence of highly pathogenic Trypanosoma evansi strains in the Philippines has resulted in substantial losses in livestock production. In this study, we isolated T. evansi from infected-water buffaloes in the Philippines and analyzed their virulence using mice and cattle. A total of 10 strains of T. evansi were isolated. Evaluation of the virulence of each strain using mice depicted significant differences among the strains in the prepatent period, the level of parasitemia, and the survival time of the infected animals. In mice infected with the highly pathogenic T. evansi, signs of excessive inflammation such as marked splenomegaly and increase more than 6-fold in the number of leukocytes were observed at 8 days post-infection. To study the virulence of the parasite strains in cattle (which are the common T. evansi hosts in Philippines), cattle were infected with the T. evansi isolates that showed high and low virulence in mice. The rate of parasite growth and the length of the prepatent periods were found to be similar to those observed in mice for the respective strains. The cattle infected with the highly pathogenic strain developed anemia and a marked decrease in leukocyte counts. To determine the cause of the pathological changes, we analyzed the expression levels of inflammatory cytokines and observed up-regulation of tumor necrosis factor-α in anemic infected cattle. Our findings suggest that the epidemic of T. evansi in the Philippines is characterized by T. evansi strains with varying virulences from low to very high pathogenicity in cattle.

Interference between Theileria orientalis and hemotropic Mycoplasma spp. (hemoplasmas) in grazing cattle

Theileria orientalis and hemotropic Mycoplasma spp. (hemoplasmas) can cause anemia in cattle. Cattle infected with one of these two pathogens tend to resist infection by the other pathogen. This is called the “interference phenomenon”. However, the detailed investigation of this phenomenon using molecular techniques has not been performed until now. We used PCR to analyze blood samples from cattle grazing in two pastures, and investigated interference between T. orientalis and hemoplasmas. Results indicate that single infection with T. orientalis or hemoplasmas was more common than co-infections. The degree of anemia observed in co-infected animals was significantly milder compared to those only infected with T. orientalis. These findings revealed that the interaction between these two pathogens in cattle demonstrated interference.