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Item Histopathological and Molecular Characterization of Canine and Feline Neoplasms: An Integrated Approach Using Immunohistochemistry, PCR and qPCR(Faculty of Veterinary Medicine, Chattogram Veterinary and Animal Sciences University, Khulshi, Chattogram-4225, Bangladesh, 2025-07) Nova, Farnaz KaderNeoplastic conditions in companion animals are increasingly recognized in clinical veterinary practice, particularly in dogs and cats. These tumors often exhibit diverse biological behaviors, histopathological patterns and clinical outcomes, posing diagnostic and therapeutic challenges. The objective of this study was to comprehensively evaluate four clinical cases of neoplasia-two in dogs and two in cats- using an integrative diagnostic approach that combined histopathology, immunohistochemistry (IHC) and molecular techniques such as polymerase chain reaction (PCR), quantitative real-time PCR (qPCR) and partial sequencing. Here, case 1 featured an 8-year-old female German Shepherd with a chronic mammary gland mass. Histopathology showed ductal and nest-like epithelial arrangements with moderate pleomorphism and stromal desmoplasia. IHC revealed Ki-67 positivity in 13% of tumor cells and P53 expression in 5%. PCR confirmed expression of BCL2, Ki-67, MYC, MMP9 and P53. qPCR demonstrated upregulation of Ki-67, MYC, MMP9 and P53, suggesting transcriptionally active oncogenic pathways. The tumor was diagnosed as an intermediate-grade mammary gland carcinoma. Case 2 was a 9-year-old male local breed dog with a prepubic cutaneous mass. Histological features included squamous epithelial nests with keratin pearls, dyskeratosis and necrosis- typical of well differentiated squamous cell carcinoma. IHC showed 10% Ki-67 positivity and 7% P53 expression, indicative of modest proliferation and limited tumor suppressor involvement. PCR confirmed neoplastic gene expression, but qPCR was not performed. The diagnosis was cutaneous squamous cell carcinoma. Case 3 involved a 1-year-old female cat with a recurrent mammary mass post-surgery. Histopathology revealed papillary and tubular epithelial patterns with low mitotic activity and focal necrosis. IHC results showed only 1% Ki-67 positivity and <1% P53 expression. PCR confirmed the presence of neoplastic markers, but qPCR was not feasible. Despite histological low-grade features, clinical recurrence and rapid progression suggested an aggressive course, possibly due to incomplete resection or multicentric origin and possibly a low grade feline mammary gland carcinoma. Case 4 concerned a 4-year-old male cat with a nasal bridge mass and facial swelling. Histology revealed infiltrative glandular structures with marked atypia and high mitotic activity. Stromal desmoplasia and subcutaneous invasion were also observed. IHC showed 30% Ki-67 positivity, indicating high proliferation, while P53 staining was minimal (<1%). PCR detected all targeted oncogenes. qPCR revealed high expression of Ki-67, MYC and BCL2; MMP9and P53 were also elevated, though less significantly. The diagnosis was high-grade nasal adenocarcinoma. Across all cases, Ki-67 levels strongly correlated with tumor grade. High-grade tumors (Case 4) showed elevated Ki-67 protein and mRNA expression, while low-grade tumors (Cases 2 and 3) showed minimal Ki-67 positivity. MYC expression was notably upregulated in all tumors, indicating its central role in oncogenesis across species. BCL2 was more prominent in feline tumors, particularly in the nasal adenocarcinoma, suggesting species-specific apoptotic regulation. MMP9 was elevated in canine cases, aligning with its known role in tissue invasion and remodeling. P53 expression varied, with discrepancies between IHC and qPCR in some cases (Case 1), potentially reflecting post-translational regulation or degradation. Hematological findings supported these pathological observations. All animals showed elevated erythrocyte sedimentation rates (ESR), indicating chronic inflammation. Lymphocytosis was evident in feline cases, possibly due to prolonged antigenic stimulation from neoplastic processes. Among the amplified genes, only BCL2 produced good-quality sequence data. Several SNPs were detected within the sequenced region of BCL2, with cats showing more variants than dogs. However, as only partial mRNA and CDS regions were obtained, full-length sequencing will be required to confirm and characterize these mutations. Sequencing of other genes (Ki67, MMP9, MYC, and P53) was not successful due to poor-quality reads. This study underscores the utility of a multimodal diagnostic approach in veterinary oncology. Histopathology provided tumor classification and grading; IHC enabled assessment of cell proliferation and tumor suppressor protein involvement; PCR/qPCR offered insights into gene expression and oncogenic activity. In conclusion, integrating histological, immunohistochemical and molecular diagnostics enhances the understanding and classification of tumors in companion animals. This approach supports more precise diagnosis, better prognostication and lays the groundwork for individualized therapeutic strategies. Future research should aim to validate these molecular markers in larger cohorts and further explore their potential in targeted veterinary cancer treatments.Item Application of different diagnostic techniques in detection and estimation of prevalence of avian influenza in poultry of Chattogram region(Faculty of Veterinary Medicine, Chattogram Veterinary and Animal Sciences University, Khulshi, Chattogram-4225, Bangladesh, 2025-06) Ara, JahanAvian influenza (AI) is an infectious disease of birds caused by influenza A virus, posing a major global public health and economic concern. In Bangladesh, AI outbreaks in commercial and backyard poultry are frequent and cause huge losses in the poultry industry. This study was conducted to detect avian influenza virus (AIV) in layer poultry farms of Chattogram region by using molecular techniques and to study the pathological alterations in AIV-infected birds. A total of 108 pooled samples were collected from 26-layer farms. Gross and histopathological examinations were performed for assessing pathological changes. RNA was extracted and after cDNA conversion subjected to conventional PCR. Real-time RT-PCR was also performed targeting the M gene for AIV detection. Out of 26-layer farms, 3 were found positive for AIV based on rRT-PCR, although 6 farms were positive by conventional PCR. On the other hand, gross lesions suggestive of AIV infection were observed in birds from 8 farms (30.8%). Gross lesions included erythema and congestion in abdominal skin, hemorrhages in trachea and lungs, hemorrhage on the fat of abdomen and coronary groove of heart, enlarged spleen, and necrosis in intestine. In case of laying hen, the ova were hemorrhagic and misshapen. Histopathological changes comprised epithelial desquamation, inflammatory cell infiltration, congestion, hemorrhages, and necrosis in respiratory and digestive organs. This study confirms the AIV in layer poultry of Chattogram and provides valuable insights into the associated pathological alterations. The findings highlight the urgent need for continuous surveillance and effective control strategies against AIV to safeguard poultry health and production in Bangladesh.Item Clinicopathological and Molecular Insights into Reproductive Disorder in Dairy Cows: Hematobiochemical, Hormonal and FOXP3 Genotypic Correlations(Faculty of Veterinary Medicine, Chattogram Veterinary and Animal Sciences University, Khulshi, Chattogram-4225, Bangladesh, 2025-06) Sultana Poly, NigarReproductive efficiency is fundamental to sustainable dairy production but is constantly challenged by multifactorial infertility disorders associated with nutrition, disease, hormonal balance, and genetics. Therefore, this study aims to investigate hematobiochemical, hormonal, and genetic factors underlying cows with reproductive disorders compared to fertile cows. A case control study was conducted from June 2024 to June 2025 across three dairy-producing regions of Bangladesh (Rangpur, Bogura, and Chattogram) to identify hematobiochemical, hormonal, and genetic factors associated with reproductive disorders in dairy cows. Blood samples and fertility data were collected from 97 lactating cows from both commercial farms and smallholder systems classified into control group (conceived ≤2 AIs; n = 57) and case group (≥3 AIs or repeat breeding; n = 40). Hematological indices, serum biochemical markers, and hormonal profiles were measured, alongside molecular genotyping of some fertility-associated genes (APOB, TFB1M, FOXP3, and IARS) using conventional PCR, and TaqMan probe based real-time PCR genotyping assay, with selected confirmation by Sanger sequencing. Statistical analyses were performed using R for non-parametric tests. The results revealed significant alterations in hematobiochemical profiles in case group compared with control group counterparts although some values present within the reference range. Hemoglobin concentrations were significantly lower in case group (9.65 ± 0.93 g/dL) than control group (10.47 ± 1.03 g/dL; p = 0.002), indicating mild anemia. Glucose, total protein, albumin, and calcium, were significantly reduced in case group, suggesting poor nutritional status, altered protein metabolism, and impaired immune response. Conversely, hepatic enzyme activities (ALT and AST) were significantly elevated in case group, indicating possible hepatocellular stress or subclinical hepatic dysfunction. Lipid profiles also revealed marginally elevated LDL in case group, while HDL and triglycerides did not differ significantly. We found a clear association between endocrine imbalance and reproductive inefficiency. Fertile cows had significantly higher serum FSH (3.72 ± 2.08 mIU/ml vs. 1.90 ± 2.41 mIU/ml; p < 0.001), progesterone (3.28 ± 1.40 ng/ml vs. 2.47 ± 1.76 ng/ml; p = 0.01), and AMH (0.20 ± 0.12 ng/ml vs. 0.05 ± 0.04 ng/ml; p = 0.002) compared to case group. These findings suggest that insufficient gonadotropin stimulation, reduced ovarian reserve, and luteal insufficiency contribute to infertility in dairy cows. Cows under poor management and nutrition also exhibited significantly lower Hb, glucose, albumin, and calcium, as well as higher ALT and LDL, compared with cows on balanced diets. This highlights the combined role of inadequate feeding and metabolic derangements in exacerbating reproductive failure. However, real-time PCR identified the presence of allelic variations at the FOXP3 locus (g.87298881A>G) which is recently reported for recurrent infertility in different breeds of cattle except Bangladeshi cows. In our study we identified G allele frequencies ranged from 0.375 to 0.517, with moderate to high prevalence across 97 cows of Jersey, Deshi, Holstein-cross, and Sahiwal breeds. Importantly, the G/G genotype also significantly associated with reduced hemoglobin, lower glucose, higher ALT and CRP, and prolonged calving intervals, days open, and service per conception, indicating a strong correlation between the FOXP3 variant and reproductive inefficiency. Genotypic stratification also revealed that cows with the wild (A/A) and carrier type (A/G) genotype of FOXP3 gene exhibited higher FSH and progesterone levels than those with mutant type (G/G) genotypes, implicating this locus might have pathogenic role in reproductive function regulation. Genetic screening also conducted for APOB gene (1.3kb insertion mutation) for Cholesterol deficiency (CD), TFB1M gene (138 kb deletion mutation) for lethal haplotype 5 (HH5), and IARS gene mutation (c.235G>C) for weak calf syndrome and calf mortality but no carrier or mutant animals found in this cohort. Overall, this study provides the first integrated evidence in Bangladesh linking metabolic dysfunction, hormonal deficits, and the FOXP3 risk allele to poor reproductive performance related to infertility in dairy cows. These findings support integrating routine biomarker screening with genetic testing can enable early identification of at-risk animals, guiding targeted nutritional and breeding interventions to enhance fertility and sustainable productivity.Item Molecular Identification of Tick-Borne Pathogens from Tick Samples of Cattle in Bandarban District, Bangladesh(Faculty of Veterinary Medicine Chattogram Veterinary and Animal Sciences University Chattogram – 4225, Bangladesh, 2024-06) Khatun, Mst. MituThe role of tick-borne pathogen transmission is exacerbated by international cattle trade, leading to significant economic losses through diseases like anaplasmosis, babesiosis, and theileriosis. These diseases hinder cattle health, growth, and production. A cross-sectional study was conducted in the Bandarban district of Bangladesh to identify tick-borne pathogens in cattle. The study aimed to morphologically identify ticks and molecularly identify tick-borne diseases from tick samples. Tick samples (130) were collected from three upazilas: 48 from Bandarban sadar, 44 from Lama, and 38 from Alikadam along with details on each cattle's age, sex, species, and physiological status. Morphological identification under a microscope revealed that Boophilus sp. and Haemaphysalis sp. had an overall prevalence of 92.31% and 7.69%, respectively. Tick infestations were more common in the summer (94.44%) compared to spring (87.5%). Adult cattle showed higher susceptibility to Haemaphysalis sp. (13.33%) compared to young cattle and calves (0%), while Boophilus sp. was more prevalent in young cattle and calves (100%) compared to adults (86.7%). Indigenous breed cattle were more prone to tick infestations (95%) than crossbred cattle (75%). The prevalence of Boophilus sp. was highest in Lama (33.85%), followed by Bandarban Sadar (33.08%) and Alikadam (25.38%). Haemaphysalis sp. was found only in Bandarban Sadar (3.85%) and Alikadam (3.85%). PCR testing of 20 pooled tick samples identified tick-borne pathogens, resulting in three Anaplasma marginale positive samples (one each from Bandarban Sadar, Lama, and Alikadam), one Babesia bigemina positive sample from Alikadam, and one Theileria annulata positive sample from Bandarban Sadar. Sequencing and phylogenetic analysis revealed the presence of Anaplasma marginale, Babesia bigemina, and Theileria annulata in cattle in the Bandarban district, with these pathogens being carried by ticks. The findings of this study are invaluable for veterinarians and cattle farmers in Bandarban, providing insights necessary to develop effective prevention and control measures against tick-borne infections..Item PATHOLOGICAL AND MOLECULAR INVESTIGATION OF HEPATOMEGALY AND SPLENOMEGALY SYNDROME IN POULTRY IN CHATTOGRAM(Chattogram Veterinary and Animal Sciences University, Khulshi-Chattogram-4225, 2023-06) Islam, Md. MuhyminulEnlarged liver and spleen are notable pathological lesions in poultry indicative of many infectious diseases that leads to economic losses due to high mortality rates and reduced egg production. Various bacteria, viruses, and infectious agents are associated with these notable lesions in the poultry. In this study, we conducted a cross-sectional study using enlarged liver and spleen samples from 35 different dead chickens (15 layers and 20 broilers). These samples were collected after postmortem examination from the dead birds brought to the department of Pathology and Parasitology at Chattogram Veterinary and Animal Sciences University (CVASU), by different poultry farm owners in Chattogram district for diagnosis and treatment purposes. Among those samples 4 different layer birds had whitish nodular lesions in the liver and spleen and tentatively diagnosed as Marek’s disease (MD) or Lymphoid leukosis (LL). However, in other cases congestive lesions were markedly observed in liver and spleen and were diagnosed as E. coli, Salmonella or other viral infections. There were no perihepatitis or hemorrhage in abdominal cavity in any cases. After necropsy, swab samples from liver and spleen were initially inoculated into selenite cystine broth and blood agar, and subsequently streaked onto EMB and XLD agar for the isolation and identification of bacterial colony. A part of the samples were also preserved in 10% buffered formalin for histopathological examinations, and in -200C for molecular work. After bacteriological culture 17 (48.5%) samples showed predominantly positive results for E.coli on EMB agar, while no Salmonella sp were positive on XLD agar, based on colony characteristics. For molecular confirmation DNA was extracted from those 4-layer bird samples (liver and spleen) that were suspected MD/LL, and PCR was done initially using ICP4 gene primers for MD virus. Finally, it’s found that 3 samples were amplified for ICP4 gene and confirmed to be Marek's disease. But due to limitations RNA couldn’t be extracted and RT-PCR couldn’t be done to check another sample with nodular lesion which might be LL or due to other RNA viruses. Furthermore, the liver and spleen samples with nodular lesions and some other samples were investigated for histopathological lesions which reveals notable findings include- severe congestion and hemorrhages, neoplastic foci with the accumulation of pleomorphic cells specially lymphoblast, various sized lymphocyte, and macrophages in the liver. In spleen, various sized lymphocytes along with extended white pulp also found which is composed of reticular cells indicative xii for extensive inflammatory reaction. In conclusion, molecular diagnosis is necessary for hepatomegaly and splenomegaly along with the histopathological examinations that is associated with any lymphoproliferative lesions.Item Pathological investigations of disease conditions in broiler chickens predisposed to dust in dry(Chattogram Veterinary and Animal Sciences University, Khulshi-Chattogram-4225, 2023-06) Paul, SubrataDust is a common pollutant present in the air of poultry houses. These dust particles has a great impact on the health of the birds. This study aims to find out the pathological effect of dust in chickens. In this study, the pathological investigation was carried out by studying tissue samples taken from dead birds with the history of using sawdust as litter material in poultry houses during the dry season. A total of 168 samples were collected from 83 affected birds over the period of November 2021 to April 2022. Gross pathological findings include congested trachea, lungs, and liver. Ascites is also observed in most cases. Microscopic examination revealed congestion, hemorrhage, infiltration of inflammatory cells, deciliation, and desquamation of lining epithelia in the trachea. The presence of dust particles surrounding the alveolar lining and metaplasia of alveolar lining epithelia was observed in lung sections with severe congestion, inflammatory cellular infiltration, emphysema, and pulmonary edema. Coagulation necrosis and marked congestion around the central vein were identified in liver tissues. In conclusion, the respiratory system of poultry is irritated by dust, leading to pulmonary hypertension and ascites, which can cause severe discomfort and pain, and may result in death.Item Histopathological Investigation of Selective Neoplasm Found in Dog and Cat(Chattogram Veterinary and Animal Sciences University, Khulshi-Chattogram-4225, 2023-06) Dhar, AnamikaHistopathological investigation of tumor cells plays a pivotal role in the diagnosis, prognosis, and treatment of various neoplastic conditions in companion animals. This study aims to explore the diverse cellular characteristics of suspected tumor cases includes lipoma in a cat, and a case of lymphoma and mammary gland carcinoma in two individual dogs using histopathological examinations. The tissue samples were collected after the surgical removal of the tumors from the respective animals that were documented at the Shahedul Alam Quadary Teaching Veterinary Hospital (SAQTVH) in Chattogram and the Teaching and Training Pet Hospital and Research Center (TTPHRC) in Dhaka. Grossly a large nodular mass were palpated in intra-abdominal region in a 5 years old local spayed cat and laparotomy was performed at TTPHRC, revealed a yellow, soft, lobulated mass with an oily surface. Besides, a 1.2 years old male Rottweiler dog with an around 5 cm nodular mass in left submandibular region and a local 8 years old unspayed dog with enlarged masses in mammary gland region were admitted in SAQTVH and TTPHRC respectively for surgery. Histopathological examination revealed mature adipocytes with eccentrically placed nuclei and less mitotic figures that indicate benign form of neoplasm in the cat called lipoma. In the Rottweiler dog the tumor tissue showed large number of pleomorphic cells, anisokaryosis, uneven distribution of chromatin materials, increased amount of binucleated cells indicating increased mitotic figure and fibroblast which is characteristics figure of lymphoma. The neoplastic cells also present within the blood vessels suggestive of metastasis and poor prognosis. In mammary gland tumor the ductal epithelial cells were proliferated, resulting in the creation of finger-like projections that extend into the inner space (lumen) and presence of pleomorphic cells with metaplasia within the milk duct indicative of invasive ductal carcinoma in dog. In Bangladesh, very few studies have been conducted on tumor in companion animals. Therefore, it‘s essential to enhance our understanding of tumor biology and providing clinicians with valuable information to make perfect decisions in tumor management. Detailed investigation on the histopathology of abnormal cellular growth in this study will be helpful for diagnosis, prognosis and treatment of such cases in dogs and cats.Item Pathological Investigation on different forms of E. coli infections (colibacillosis) in broiler chicken and molecular confirmation(Chattogram Veterinary and Animal Sciences University, Khulshi-Chattogram-4225, 2023-06) Jahan, SadiaAvian colibacillosis is a significant global infectious disease in birds of all ages, leading to substantial economic losses in poultry production. This disease is caused by Escherichia coli infection which is a highly prevalent bacterial pathogen in man and animals. Colibacillosis is a multifaceted syndrome distinguished by various organ abnormalities, including air sacculitis (61.67%), pericarditis (58.33%), perihepatitis (33.33%), yolk sac infection (35%), enteritis (31.67%). Research involving pathological examinations was conducted on spontaneous occurrences of avian colibacillosis to investigate the prevalence and pathological manifestations of Escherichia coli infection. After necropsy, we collected samples from lung, liver and spleen for bacterial isolation and subsequently for histopathological examinations. Following the bacterial isolation process, when we cultured the bacteria on agar, the outcomes revealed colonies with a distinct pink coloration on MacConkey agar and a distinctive metallic sheen on EMB agar. These characteristics led to the identification and consideration of E. coli in 45 cases. To confirm the presence of E. coli, PCR was conducted using the housekeeping gene primer (adk), resulting in the confirmation of E. coli in 37 suspected cases initially identified from agar cultures. Microscopic examinations revealed severe congestion (83.33%), hemorrhage (90%), focal area of necrosis (76.67%), prominent nuclear and cytoplasmic changes in hepatocytes with like fatty change (33.33%) in liver. In lung, edema (20%) and congestion (75%) were commonly observed almost all cases with thickening of alveolar septa. Hemorrhage and congestion (66.67%) also observed commonly in spleen along with depletion of lymphoid elements and multiple focal areas of necrosis (33.33%) in the primary follicles resulted in the formation of secondary germinal centers. Depletion of lymphocytes at times also accompanied by reticular cell proliferation (50%) in spleen. This study underscores the significant cellular-level changes associated with colibacillosis in commercially raised broiler chickens and suggests the need for further investigation to identify the specific serotypes of E. coli present in these chicken populations.Item PATHOLOGICAL INVESTIGATION AND MOLECULAR IDENTIFICATION OF INFECTIOUS BURSAL DISEASE IN BROILER CHICKENS(Chattogram Veterinary and Animal Sciences University, Khulshi-Chattogram-4225, 2023-06) Habiba, Umme AyshaInfectious Bursal Disease (IBD) represents a highly contagious viral illness affecting young chickens, leading to immunosuppression, increased mortality rates, and stunted growth. The widespread impact of this disease has significantly undermined the poultry industry. This cross-sectional study aimed to explore the gross and histopathological alterations within various organs in suspected cases of IBD, subsequently confirmed through reverse transcription (RT-PCR) for molecular validation. This study was conducted at the Pathology Laboratory of Chattogram Veterinary and Animal Sciences University from December 2021 to February 2023. Out of 200 suspected cases of IBD identified based on gross lesions, 30 cases (comprising bursa, kidney, and spleen samples from each case) were randomly selected for histopathological investigation, while 8 samples among them were chosen specifically for molecular diagnosis. The clinical signs observed in IBD infected chickens were whitish diarrhea mixed with blood, ruffled feather, and massive death within short period. Necropsy findings revealed petechial hemorrhage in the bursa of fabricius, kidneys, thymus, spleen, and thigh muscle. Over the disease course, the bursa of fabricius displayed edematous changes in its serosal and mucosal regions, transitioning to a whitish creamy appearance and subsequent atrophy. The kidneys exhibited paleness, edema, and hemorrhagic manifestations. Depletion of follicular lymphocytes as well as edema, formation of cortical rim, interfollicular hemorrhage were the major histopathological changes found in bursa of fabricius and spleen. Kidney sections revealed hemorrhage, congestion, infiltration of inflammatory cells between renal tubules, edema in renal tubules, partially detached lining epithelium from basement membrane of many tubules. Out of thirty clinically diagnosed samples, eight samples were selected and all detected positive by RT-PCR for the hyper variable region of VP2 gene. The consistent correlation among these diagnostic modalities strengthens the confidence in utilizing gross lesion assessments, supported by histopathological and molecular analyses, as effective and reliable means for identifying Infectious Bursal Disease Virus (IBDV) infected chicken.Item Molecular identification of babesiosis in stray dog at Chittagong Metropolitan Area(A thesis submitted in the partial fulfillment of the requirements for the degree of Master of Science in Parasitology Department of Pathology and Parasitological Faculty of Veterinary Medicine Chittagong Veterinary and Animal Sciences University Chittagong-4225, Bangladesh, 2014-12) E-Azam, Md. NurAn epidemiological study of babesiosis in stray dogs was conducted at Chittagong Metropolitan area, Bangladesh, for six months. Blood samples of 130 stray dogs were collected from nine randomly selected locations along with the information of age and sex. Blood samples were initially examined by Giemsa's stained blood smear method. DNA was extracted from microscopy positive blood samples and Babesia spp were confirmed by amplifying 18S rRNA gene. The prevalence was determined 6.92% in microscopic technique and the percentages of PCR positive samples from the microscopy positive samples were 66.67% (6 out of 9). The highest prevalence of babesiosis was found in Alankar (16.67%) compared to other areas. Significantly higher frequency of babesiosis was found in adult dog than younger (p<0.05). Again, Babesia infection was common in male (11.94%) than female (1.59%) and were statistically significant (p<0.05). Furthermore, we sought to develop a seminested PCR to detect and differentiate Babesia gibsoni (Asia genotype), B. canis subsp. vogeli, B. canis subsp. canis and B. canis subsp. rossi DNA in canine blood samples. An outer primer pair was designed to amplify a 340 bp fragment of the 18S rRNA genes in the seminested primary reaction. Then specific internal primers were designed for B. gibsoni (Asian type) (BgibAsia-F) and B. canis subsp. canis (BCC-F) that were paired with the outer reverse primer in the seminested secondary reaction to amplify 185 bp and 198 bp respectively. Based on seminested PCR and sequencing of the PCR products, all parasitic isolates were identified as Babesia gibsoni (Asian type) and B. canis subsp. canis. These results recommended that Babesia gibsoni (Asian type) and B. canis subsp. canis parasites are widespread in stray dog in Chittagong Metropolitan area, Bangladesh. Further investigations are required to clarify the origin, distribution, vector and pathogenesis of these parasites tracking in dogs in Bangladesh.
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