<div style="text-align: justify;">This study was designed to determine the prevalence and endemic status of Anaplasma marginale in cattle in the province of Konya. A total of 700 cattle were randomly selected, categorized in different age groups (i.e. 0&ndash;6, 7&ndash;12, 13&ndash;24, 25&ndash;36 and &gt;36 months) and examined in Kadinhani, Cumra and Beysehir provinces and in Konya city center. The presence of A. marginale was diagnosed by microscopic examination of blood smears and competitive ELISA (cELISA). Parasites were observed in 11.29% of blood smears, and 31.86% of animals were positive for antibodies against A. marginale. The rates of seropositivity in successively older age groups were 19.1, 24.39, 33.14, 44.21 and 37.36% respectively. The endemic status of the disease was determined by calculating the inoculation rate (h) of cattle in each age group. The h value was detected to be lower than 0.005 and the endemic status of A. marginale was found to be unstable. If the cattle in Konya province were vaccinated when they are 9 to 12 months of age, they could be protected during the seasons when the disease is prevalent.</div>

The endemic status of Anaplasma marginale in cattle, in Turkey

<div>Background: Bovine anaplasmosis caused by Anaplasma marginale is highly endemic in tropical and sub-tropical regions of the world where it accounts for huge economic losses to the livestock sector. Approximately 30% of water buffalo (Bubalus bubalis) herds in India are located in Uttar Pradesh (U.P). However in India, most of the research studies on Anaplasma are focused on cattle whereas studies on this pathogen in water buffaloes have been neglected. Therefore, present investigation was undertaken for determining the prevalence of A. marginale infection in water buffaloes in U.P, North India.&nbsp; 
 
Methods: In the current investigation during 2017-2019, the prevalence of A. marginale infection in 228 water buffaloes in three districts of North-West U.P was studied by microscopy and by nested-PCR amplification of major surface protein 5 (msp5) gene.&nbsp; 
 
Result: Microscopic examination of blood smears of 228 water buffaloes revealed 7.89% animals positive for A. marginale. Conventional PCR of full length major surface protein 5 (msp5) gene detected 29.39% and nested-PCR amplification of this gene detected 64.0% of these animals as positive for A. marginale infection. A high incidence of the infection detected in this study area suggests that water buffaloes may serve as reservoir hosts of A. marginale and play a vital role in the spread of infection to other susceptible animals. However further studies are required on larger buffalo populations for determining the epidemiological status of anaplasmosis in these animals in India.</div>

Prevalence of Anaplasma marginale in Water Buffaloes (Bubalus bubalis) in the State of Uttar Pradesh, North India

Background: Bovines are domesticated ungulates mainly raised as livestock for milk, meat and leather and as draught animals. A slaughterhouse study helps to assess the disease status of herds and contains valuable information about the incidence and epidemiology of animal diseases. Haemoprotozoan diseases are one of the major problems which adversely affect the health and productivity of cattle. The diverse climatic zones of India are highly conducive to the survival and propagation of vectors and vector-borne pathogens. However, there is no data on proper slaughterhouse study of bovines in Mizoram even in India for determination of the prevalence of haemoprotozoan infections. 
 
Methods: The blood samples were collected in an EDTA vials. The blood smear was prepared, stained with Giemsa stain and examined for the presence of haemoprotozoan. The DNA was isolated from positive samples by use of a DNeasy Blood and Tissue Kit. Then PCR was performed for all the isolated DNA collected from positive samples. After confirmation through PCR, the representative samples for Anaplasma sp. and Theileria sp. were sent for sequencing. Then the sequencing results were annotated using BLAST, ClustalW multiple alignment program of MEGA9 software. The phylogenetic tree was generated by using the Neighbour-Joining method, keeping bootstrap consensus from 1000 replicates.&nbsp; 
 
Result: After the blood smear examination, PCR was performed for confirmation. Molecular detection of Anaplasma marginale, Theileria orientalis was done. Molecular characterization of 2 isolates from Anaplasma marginale and Theileria orientalis was performed by sequencing. The sequence analysis of rpoB gene of isolates of Anaplasma revealed both the isolates were placed in the same clade in the phylogenetic tree. The phylogenetic analysis of Thileria isolates revealed that isolate Miz-86 was closely placed with the Type-N3 clade. Whereas the other isolate i.e. Miz-87, belonged to the Type-5 clade.

Molecular Characterisation of Anaplasma marginale and Theileria orientalis in Slaughtered Bovines of Aizawl District, Mizoram

Background: This study aims to identify risk factors for Rhipicephalus (Boophilus) microplus ticks&rsquo; infestation in South Gujarat&rsquo;s cattle and explore its implications on Anaplasma marginale. 
 
Methods: An approach was employed to retrieve the crucial insights from the owners and examinations of 2305 cattle (crossbred=1495, indigenous=810) of South Gujarat, India. Species specific PCR was used to detect 576 bp of msp5 gene of A. marginale in the oviposited ticks.&nbsp; 
 
Result: Crossbred cattle (73.91%) had higher tick prevalence than indigenous ones (55.80%), with R. (B.) microplus being the dominant species. Highest tick infestation was noted in cattle &gt;3 years (74.43%), followed by &lt;1 year (70.23%) and 1-3 years old (55.60%). Ticks collected from crossbred exhibited higher prevalence of A. marginale compared to indigenous cattle. Significantly highest (p &gt;0.05) ticks infestation rate was noted in cattle with pale conjunctiva, poor body condition and black coat colour. Infection rate of both the parasites was highest during summer, followed by monsoon and winter. Kachcha house and earthen floor favoured the survival of the pathogen. Owners in surveyed regions combat tick infestations using mechanical (7.69%), chemical (83.51%) and herbal/no treatments (10.07%). These findings emphasize the intricate link between tick infestations and A. marginale infection, offering crucial insights for cattle health management.

Rhipicephalus (Boophilus) microplus Infestation and its Implications in Anaplasma marginale Infection among Cattle in South Gujarat, India

<div>Background: Bovine anaplasmosis is a haemolytic disease of cattle caused by an obligate intra-erythrocytic bacterium Anaplasma marginale and is characterized by high fever, dyspnoea, anaemia, icterus, decreased milk production and death. The present study was undertaken to study the gross and histopathological changes caused by naturally occurring A. marginale infection in cattle.&nbsp; 
 
Methods: History and clinical examination of animals revealed inappetance/anorexia, presence of tick infestation, high fever, pale or icteric mucous membranes, tachycardia, dyspnoea and lymphadenomegaly. The diagnosis of A. marginale infection was done by blood smear examination and further confirmed by polymerase chain reaction and sequencing analysis. During the present study, two animals did not respond to treatment and were succumbed to A. marginale infection. A systematic post-mortem examination was performed on the animals and gross lesions were recorded. The tissue samples from various vital organs such as heart, lungs, liver, spleen and kidneys were collected in 10% neutral buffered formalin and processed for histopathological examination.&nbsp; 
 
Result: The post-mortem examination revealed emaciated carcass with yellowish discoloration of serosal surfaces of the abdominal organs, haemorrhagic heart, congestion of lungs, hepatomegaly, distension of the gallbladder and splenomegaly. The major histopathological changes noticed included myocardial degeneration and necrosis, interstitial pneumonic changes, degenerative changes in hepatocytes and renal tubular epithelial cells, enlargement of red pulp area of spleen with histiocytic proliferation and hemosiderosis. Histopathology also revealed inflammatory process characterized by infiltration of mononuclear cells in various organs. Very few reports are available on the pathological aspects of A. marginale infection and thus the outcome of the present study can yield valuable information to aid in the diagnosis of bovine anaplasmosis under field conditions.</div>

<div>Gross and Histopathological Findings of Naturally Occurring Anaplasma marginale Infection in Cattle</div>

<div>Background: A novel, rapid and specific multiplex polymerase chain reaction was developed to diagnose hemo-parasitic infection in bovine blood co-infected with three of the most common hemo-parasites.&nbsp; 
 
Methods: The diagnostic process relied on the detection of the three different bovine hemoparasites isolated from red blood cells (RBCs) of cattle (N=30) by conventional Giemsa stained blood smear (GSBS) and confirmed by multiplex PCR. The multiplex PCR system was used to diagnose GSBS positive blood samples (N=12) found infected or co-infected with hemoparasites. The designed multiplex primer sets was attempted to amplify 205, 313 and 422 bp fragments of apocytochrome b, sporozoite and macroschizont 2 (spm2) and 16S rRNA gene for Babesia bigemina, Theileria annulata and Anaplasma marginale, respectively. 
 
Result: This multiplex PCR was sensitive with the ability to detect the presence of 150 ng of genomic DNA. The primers used in this multiplex PCR also showed highly specific amplification of specific gene fragments of each respective parasite. Comparing the two detection methods revealed that 58.33% of specimens showed concordant diagnoses with both techniques. The specificity, positive predictive value and kappa coefficient of the agreement was highest for diagnosis of B. bigemina and lowest for A. marginale. The overall Kappa coefficient for diagnosis based on GSBS for multiple pathogens compared to multiplex PCR was 0.56, slightly behind the threshold of 0.6 of agreement. Therefore, confirmation should always be based on PCR to rule out false positives due to differences in subjective observations, stain particles and false negatives due to low parasitemia. The simplicity and rapidity of this specific multiplex PCR method make it suitable for large-scale epidemiological studies and follow-up of drug treatments.</div>

<div style="text-align:justify">&nbsp;</div>

Development of Novel Multiplex PCR for Diagnosis of Co-infected Hemo-parasites in Cattle

A crossbred cow aged 6 year was presented to Teaching Veterinary Hospital of the Institute with the history of fever, tick infestation, anemia and decreased milk yield since last 10 days. Clinical examination revealed fever, pale mucous membrane and nil rumen motility. Hematological examination findings were anemia, leucopenia, relative neutrophilia and lymphopenia and decreased platelets. Blood smear examination revealed Anaplasma Marginale infection. Animal was treated with blood transfusion, oxytetracyclin, oral hematinic and multivitamin. After half an hour of start of transfusion, cow showed transfusion reaction in the form of urticarial eruptions all over the body, edema of eyelids, lips and over the face, frothy discharge from mouth with muscle tremor and shivering. Transfusion reaction was successfully managed with Adrenalin, Dexona, Avil along with normal saline.&nbsp;

Therapeutic management of blood transfusion reaction in a crossbred cow

Background: Haemoparasitic diseases in cattle are infectious conditions primarily transmitted by arthropods, particularly ticks. In India, especially in the northeastern region (NER), these diseases lead to significant economic losses for farmers, impacting both production and treatment costs. This study aims to investigate the prevalence of haemoparasitic diseases in cattle around Guwahati, as there is a lack of comprehensive data on their prevalence over an extended period in this area. 
&nbsp; 
Methods: The study was conducted for a period of five years from 2017 to 2021. To study the prevalence of haemoparasite Giemsa-stained blood samples from cattle presented to OPD of Veterinary Clinical Complex, CVSc, AAU, Khanapara were examined. The study was carried out concerning breed, age and sex of the animal and also regarding the prevailing seasonal changes. 
&nbsp; 
Result: A total of 2100 cattle were screened, out of which 1767 (84.14%) were found to be infected with haemoparasites. In the present study, infection with Theileria was found to be the highest (72.94%) followed by Anaplasma marginale (12.00%) and Babesia bigemina (11.09%). A mixed infection of haemoparasite was also recorded as 3.96%. The parasites were identified based on morphology in Giemsa-stained blood smear. The highest infection was recorded in the age group of &gt;5 years (89.23%) followed by 1-5 year group (85.99%). Similarly female animals are found to be more infected (84.92%) than that of the male (77.14%). Seasonally, the prevalence of haemoprotozoa was found highest in monsoon (97.38%) and lowest in winter (46.13%). The present study provides systematic data on haemoparasite infection in cattle.

Temporal Trends of Tick-borne Haemoparasitic Infections in Cattle of Assam, India: A Five-year Retrospective Study

A total of 480 suspected blood samples were collected from cattle (n=254) and buffaloes (n=226) during the study period from September 2011 to December 2014 for determination of prevalence of haemoprotozoan parasites from South Western Gujarat (Junagadh). The conventional optical microscopy of Giemsa stained blood smears revealed that 37% of cattle and 38.93% of buffaloes were infected with haemoprotozoan parasites including Anaplasma marginale, Babesia bigemina, Trypanosoma evansi and Theileria annulata. The animals infected with haemoprotozoan diseases showed lower total erythrocyte count (TEC), haemoglobin (Hb), packed cell volume (PCV), mean corpuscular haemoglobin concentration (MCHC) and higher total leukocyte count (TLC) and mean corpuscular volume (MCV). Incidence of haemoprotozoan infection between the breeds differed significantly (P&lt;0.05). Haemoprotozoan infections were recorded significantly higher (P&lt;0.001) in monsoon season followed by summer and being lowest in winter season in cattle. Over all prevalence in female animals were higher than male counterparts. In both the species, over all higher prevalence of babesiosis was recorded in comparison to other haemoprotozoan diseases and the difference being statistically non-significant (p&gt;0.05). The prevalence of B. bigemina in bovines was associated with various risk factors namely age, season and breed (p&pound;0.05). Sex wise females recorded apparently higher infection rates than males. The multivariate logistic regression models showed that the risk of babesiosis was significantly higher in rainy season (OR=5.18, P=0.003) followed by summer (OR=3.9, P=0.019) compared to winter season. Conversely, in buffaloes, the risk of babesiosis was significantly higher in summer season (OR=9.0, P=0.004) followed by rainy (OR=7.43, P=0.008) compared to winter season. The risk for anaplasmosis in buffaloes increased by 3.46 times (OR=3.64, p=0.027) in non descriptive breed compared to well defined breed.&nbsp;

Prevalence and assessment of risk factors for haemoprotozoan infections&nbsp;in cattle and buffaloes of South-West Gujarat, India&nbsp;

Background: Anaemia is a common and important clinical presentation in goats due to infectious, traumatic, nutritional, haemoprotozoal, endo and ecto parasitic causes. Haemoprotozoan are obligate intracellular pathogens transmitted by ticks with an impact on animal health. Anaplasma ovis, Babesia ovis, Babesia motasi and Theileria luwenshuni that infects goats can be diagnosed by different methods like giemsa staining, PCR or competitive ELISA. 
 
Methods: Goats presented to Large Animal Clinics of Madras Veterinary College Teaching Hospital, Chennai, India, were screened for the signs of anaemia. In this study, PCR method was used for the detection of Anaplasma, Babesia and Theileria infection. 
 
Result: Among 25 blood samples collected from anaemic goats, 48 per cent were found positive for haemoprotozoan disease. Out of which 32 per cent were Anaplasma ovis followed by 4 per cent of Babesia motasi and Babesia ovis each whereas remaining 8 per cent showed mixed infection. This indicated a high incidence of haemoprotozoan infection as a major cause of anaemia in goats reared under field conditions.