Analysing the Impact of Resistant Starch Formation in Basmati Rice Products: Exploring Associations with Blood Glucose and Lipid Profiles across Various Cooking and Storage Conditions In Vivo.

Common cooking methods were used to prepare basmati rice products, including boiling 1 (boiling by absorption), boiling 2 (boiling in extra amount of water), frying, and pressure cooking. The cooked rice was held at various temperatures and times as follows: it was made fresh (T1), kept at room temperature (20-22 °C) for 24 h (T2), kept at 4 °C for 24 h (T3), and then reheated after being kept at 4 °C for 24 h (T4). The proximate composition, total dietary fibre, resistant starch (RS), and in vitro starch digestion rate of products were examined. The effect of RS on blood glucose and lipid profiles was measured in humans and rats, including a histopathological study of the liver and pancreas in rats. The basmati rice that was prepared via boiling 1 and stored with T3 was found to be low in glycaemic index and glycaemic load, and to be high in resistant starch. Similarly, in rats, the blood glucose level, cholesterol, triglycerides, and LDL were reduced by about 29.7%, 37.9%, 31.3%, and 30.5%, respectively, after the consumption of basmati rice that was prepared via boiling 1 and stored with T3. Awareness should be raised among people about the health benefits of resistant starch consumption and the right way of cooking.

Effect of cooking and storage temperature on resistant starch in commonly consumed Indian wheat products and its effect upon blood glucose level.

Background/objectives: The health benefits provided by resistant starch have been well documented; however, few studies are available on the resistant starch content of wheat products in India. Moreover, few studies have examined the in vivo efficacy of resistant starch in wheat products in improving glucose levels. This study was conducted to evaluate the effect of cooking and storage temperature on the formation of resistant starch in Indian wheat products and its effect on blood glucose levels in humans and rats. Methods: Wheat products were prepared by common cooking methods including roasting (Chapati), boiling (Dalia), Shallow frying (Paratha), and Deep frying (Poori). They were then stored at different temperatures including freshly prepared within 1 h (T1), stored for 24 h at room temperature (20-22°C) (T2), kept at 4°C for 24 h (T3) and reheated after storing at 4°C for 24 h (T4). The products were then analyzed for proximate composition (moisture, crude protein, crude fat, ash crude fibre, and carbohydrates). The effect of different cooking methods and storage temperatures on Resistant, non-resistant and total starch, total dietary fibre (soluble and insoluble), in vitro starch digestion rate (rapidly and slowly digestible starch), amylose and amylopectin content were analysed using standard operating procedures. The effect of products found to have higher resistant starch was studied on the post prandial blood glucose response of 10 healthy individuals using change in by analysing their glycemic index and glycemic load of wheat products. Further, the effect of resistant starch rich chapati on the blood glucose level of rats was also studied. Tukey's test in factorial CRD was used to assess the effect of cooking and temperature on various parameters. Results: The amount of resistant starch was found to be high in dalia (boiling, 7.74%), followed by parantha (shallow frying, 4.94%), chapati (roasting, 2.77%) and poori (deep frying 2.47%). Under different storage temperatures, it was found high in products stored at 4°C (T3), followed by products stored at room temperature (T2), reheated products (T4) and lesser in freshly prepared products (T1). The glycemic index and glycemic load were found low in chapati (43, 32.3) and dalia (41.1, 28.6) stored at 4°C (T3) compared to others. The resistant starch content found in chapati stored at T3 was found to be more effective at reducing blood glucose levels in rats from 291.0 mg/100 mL to 225.2 mg/100 mL in 28 days of study compared to freshly prepared chapati (T1) and stored at room temperature (T2). Conclusion: Cooking methods including boiling, roasting and shallow frying increased the amount of resistant starch in foods, but cooking methods such as deep frying decreased the amount of resistant starch in food. Products stored at 4°C and at room temperature for 24 h increased the amount of resistant starch whereas the products that were freshly cooked and reheated decreased the amount of resistant starch in foods. At 4°C the stored products have a high amount of insoluble dietary fibre, slowly digestible starch, high amylose and low glycemic index. They take time to digest, meaning that they slowly increase blood glucose levels. The effect of insoluble dietary fibre and resistant starch in the inhibition of glucose diffusion in the small intestine is suggested to be due to the absorption or inclusion of the smaller sugar molecules. In vivo research showed that fibre and resistant starch in the digestive system of rats acts as the main factors in slowing glucose absorption and reducing a rise in blood glucose levels by promoting glycogen synthesis and inhibition of gluconeogenesis.

Targeting the nervous system of the parasitic worm, Haemonchus contortus with quercetin.

Prevalence of infection, limited choice of drugs, and emerging resistance against contemporary medications lead to a pressing need to develop new anthelmintic drugs and drug targets. However, little understanding of worms' physiology has substantially delayed the process. Here, we are reporting the tissue morphology of Haemonchus contortus, intestinal parasitic helminths found in small ruminants, and targeting its nervous system with quercetin, a naturally occurring flavonoid. Quercetin showed anthelmintic activity against all of the developmental stages of H. contortus. Further, histological analysis demonstrated damage to various body parts, including isthmus, brut, pseudocoele, and other organs. Mechanistic studies revealed the generation of oxidative stress and alterations in the activities of the stress response enzymes, such as catalase, superoxide dismutase, and glutathione peroxidase. Moreover, the time-dependent imaging of reactive oxygen species (ROS) generated due to quercetin treatment disclosed neuropils as the primary targets of quercetin in adult worms, which eventually lead to the paralysis and death of the worms. Thus, this work demonstrates that the nervous system of the parasitic helminth, H. contortus, is a novel target of the drug quercetin.

Molecular epidemiology, phylogenetic analysis and risk assessment of Anaplasma marginale from naturally infected bovines of Punjab (India).

Anaplasmosis, an infectious vector borne rickettsial disease caused by different species of Anaplasma transmitted through mechanical and biological (tick) processes has a great concern to livestock industry due to its associated economic losses. The current cross sectional comprehensive epidemiological study was conducted from August 2020 to November 2021 on 391 (277 cattle and 114 buffaloes) bovines from different districts of five agro climatic zones of Punjab state. Classical microscopic examination of Romanowsky stained blood smears showed an occurrence of 13.00%, while DNA amplification targeting major surface protein (msp5) of Anaplasma marginale revealed the 382 bp amplicon in 32.48% samples. Zone based molecular prevalence of A. marginale was highest in the Undulating zone (41.93%) and least in the Sub-Mountain zone (18.84%). The prevalence in Central plain zone, Western plain zone, and Western zone were 40.15, 30.95 and 29.91%, respectively. An overall molecular prevalence of A. marginale was 34.52, 32.75 and 20.0% in young, adults and calves, respectively, the difference being non-significant. Anaplsma marginale was more prevalent in unorganized farms (38.22%; 60/157) than organized farms (28.63%; 66/234). Risk factors analysis revealed young, female cattle at unorganized farms were more prone to anaplasmosis. Out of 127 positive samples, four samples were customed to sequencing revealed 98-99% homology with published sequences for other available global isolates. Multi-single nucleotide polymorphisms (SNP) were observed in the sequence of two samples when aligned with the reference sequence from the NCBI database (CP023731).

Assessment of risk factors associated with prevalence of gastrointestinal parasites in poultry of central plain zone of Punjab, India.

BACKGROUND AND AIM: Parasitic diseases are an important hurdle to the economy for the developing poultry industry due to their deleterious effects resulting into malnutrition, diminished feed conversion ratio, weight loss, decreased egg production, and mortality in young birds. The aim of this study was to assess the prevalence and associated risk factors of gastrointestinal (GIT) parasites in poultry farms of central plain zone of Punjab. MATERIALS AND METHODS: A total of 490 pooled droppings and 351 intact intestines of poultry from slaughterhouses from seven districts of central plain zone of Punjab state, India, were collected and analyzed from September 2016 to May 2018 by qualitative and quantitative techniques. RESULTS: An overall prevalence of GIT parasites was 38.36% with significantly (p<0.01) highest (74.1%) in Ludhiana and lowest (12.0%) in Shri Fatehgarh Sahib. The most predominant (86.2%) infection was coccidia. The birds reared under a deep litter system were having a higher (p<0.01) fecal load of helminthic eggs and coccidian oocysts (54.4%) compared to the cage system (37.5%). Infection rate was apparently more (40%) in broilers than layers (35.7%). Prevalence of GIT parasites was higher (p<0.01) in monsoon season (58.5%) and lower in summer season (24.48%). The broilers in the age group of 0-2 weeks possessed a significant higher (p<0.05) level of GIT parasitic infection (57.5%), while in case of layers, a higher infection rate (46.66%) was observed in birds between 9 and 18 weeks of age as compared in other groups. Higher (p<0.05) infection rate of GIT parasites was seen in crossbred (45.55%) birds as compared to desi birds (20.00%). CONCLUSION: The study showed that coccidiosis was the predominant infection among all GIT parasites based on fecal and intestinal tract content analysis. The risk factors associated with the prevalence of GIT parasitic infections were geographical location, deep litter system, broilers, age, crossbred breeds, and monsoon season.

Comparative epidemiology and pathophysiology of patent and latent babesiosis caused by Babesia bigemina in buffaloes and cattle from different agroclimatic zones of Punjab State, India.

To conduct comparative epidemiology of parasitologically positive (patent) and polymerase chain reaction positive (latent) cases of bovine babesiosis in Bet Region (low-lying areas adjoining Sutlej, Beas, Ravi, and Ghaggar rivers of Punjab) of diverse agroclimatic zones of Punjab state in relation to haematobiochemical parameters as patho-physiological markers, blood samples from 783 dairy animals (487 buffaloes and 296 cattle) were analysed parasitologically by Giemsa-stained blood smears (GSBS) and by molecular-based polymerase chain reaction (PCR) targeting SpeI-AvaI restriction fragment of Babesia bigemina. We ruled out the endemicity of the disease with 2.17% patent and 3.96% latent cases of B. bigemina with significantly higher prevalence (P < 0.01) in cattle than buffaloes. The spatial distribution for a guideline to local veterinary practitioners and policy-makers indicated highest number of patent and latent cases in western zone and undulating plain zone, respectively. District wise highest prevalence of patent as well as latent infection observed in SBS Nagar of undulating plain zone showed substantial agreement (Kappa value: 0.70) between the two techniques. Haematology revealed marked microcytic hyperchromic anaemia in patent animals of group I (GSBS positive; n = 17) and latent animals of group II (PCR positive; n = 14) as compared to disease-free controls (group III; n = 10). Blood urea nitrogen (BUN), creatinine and gamma-glutamyltransferase (GGT) levels were significantly increased (P < 0.05) in group I in comparison to group II and group III indicated comparative pathogenic effect of babesiosis in patent cases. Though patent cases showed higher pathogenicity of babesiosis, diagnosis of latent infection is significant as it may act as source of infection for spread to other highly prone bovines.

Cuminaldehyde induces oxidative stress-mediated physical damage and death of Haemonchus contortus.

Cuminaldehyde (CA), a monoterpenoid, preset in many plant sources including cumin, induces reactive oxygen-related damage and death in Haemonchus contortus, a parasitic worm with an LD50, values of 127.3 ±â€¯7.5, 184.5 ±â€¯12.1 and 104.1 ±â€¯7.9 µg/mL for an adult female, adult male worms (12 h) and L3 larvae, respectively (24 h). Fifty percent of inhibition of egg hatching (IC50) was obtained at 142.4 ±â€¯11.4 µg/mL after 48 h of exposure. Scanning electron microscopy revealed physical damage to the anterior and posterior ends, intestinal, ovarian, and esophageal regions of the warms on exposure to ca. The exposure of worms to CA also led to a systemic increase in reactive oxygen species (ROS) within 3 h. The better activity was seen with CA compared to standard antihelminthic drug albendazole (Alb). 74 µg/mL CA showed 2.3 fold more increase of catalase (CAT), 0.61 fold increase of superoxide dismutase (SOD), 3.3 fold increase of glutathione peroxidase (GPx) activity and 17.5 fold increase of glutathione (GSH) activity in comparison with Alb (500 µg/mL) for the same time of exposure (3 h). A firm increase of (2.9 fold) was also observed in nitric oxide synthase (NOS) activity within 12 h of exposure with CA (74 µg/mL) in comparison with Alb. Therefore the preclinical potential of CA is much higher than widely used antihelminthic drug Alb. The results open new opportunities to explore CA as a new active antihelminthic molecule.

Biomedical Evaluation of Lansium parasiticum Extract-Protected Silver Nanoparticles Against Haemonchus contortus, a Parasitic Worm.

Here we show the novel anti-helminthic potential of Lansium parasiticum aqueous extract-protected silver nanoparticles (LAgNPs) against albendazole-resistant gastrointestinal parasite Haemonchus contortus. LAgNPs showed LD50 values of 65.6 ± 32.8 nM (12 h), 139.6 ± 39.9 nM (12 h), and 64.3 ± 8.5 nM (24 h) against adult male, female, and L3 larvae, respectively. LAgNPs was also quite effective in inhibiting egg hatching, with an IC50 value of 144.4 ± 3.1 nM at 48 h of exposure. Exposure to LAgNPs generated oxidative stress and mediated physical damage in the worms' tissue. A sharp increase in reactive oxygen species and nitric oxide synthase levels was prominent due to LAgNPs' exposure. In response to oxidative stress, a sharp increase of stress-responsive enzymes' activity, like catalase, superoxide dismutase, and glutathione peroxidase activity, along with the concentration of glutathione, was observed in worm tissue, which indicated a LAgNP-responsive alteration of metabolism. The results give rise to the opportunity for the development of alternative treatment for drug-resistant parasitic worms.

First record of Toxoplasma gondii antibodiesin Royal Bengal tigers (Panthera tigris tigris) and Asiatic lions (Panthera leo persica) in India.

The purpose of this study was to detect the antibodies against Toxoplasma gondii in Royal Bengal tigers (Panthera tigris tigris), Asiatic lions (Panthera leo persica), leopards (Panthera pardus), and elephants (Elephas maximus indicus) residing in the Mahendra Chaudhury Zoological Park, in Chhatbir, Punjab (India) during winter and monsoon seasons. Using  indirect ELISA, 20 serum samples were analysed during the winter season. Results indicated that 1 lion (5%) tested seropositive, and 3 tigers and 1 lion (20%) were considered suspect. During the monsoon, 4 individuals (2 tigers and 2 lions, 20%) were seropositive, whereas only 1 tiger (5%) gave suspected results. Significantly higher globulin, creatinine, blood urea nitrogen, phosphorus, and creatine kinase values were recorded in seropositive and suspected groups. Levels of albumin, glucose, calcium, sodium, and iron decreased significantly in the seronegative group. Results from sero-testing 40 rodents trapped in and around the park depicted the presence of antibodies against Toxoplasma gondii in 1 individual. This study reveals the haemato-biochemical alterations in both seropositive and suspected wild felids for toxoplasmosis. Moreover, it provides the first serological evidence of T. gondii exposure in wild felids, notably Royal Bengal tigers and Asiatic lions, in India.

Evaluation and correlation of oxidative stress and haemato-biochemical observations in horses with natural patent and latent trypanosomosis in Punjab state of India.

The haemato-biochemical indices and oxidative stress markers in horses naturally infected with Trypanosoma evansi were evaluated by analyzing the level of these parameters between T. evansi infected (microscopically positive patent group and PCR positive latent group) and infection free horses. To compare the hemato-biochemical indices and oxidative stress indicators, horses were divided into three categories based on diagnostic test employed and positive results obtained. These included Romanowsky stained slide positive group (Group I; n = 6), PCR positive group (group II; n = 28) and negative control group (group III, n = 30), revealing parasitologically positive patent, molecular positive latent and disease free status of horses. A significant reductions in total erythrocytes count (TEC, P = 0.01), haemoglobin (Hb, P = 0.01) and packed cell volume (PCV, P = 0.04) was noticed both in group I and group II while significant neutrophilia and lymphocytopenia was observed in group I when compared to negative control group. Substantial increase in creatinine (CRTN, P = 0.032) and gamma glutamyl transferase (GGT, P = 0.012) in group I while significant decrease in glucose (GLU, P = 0.04) and iron (Fe, P = 0.01) were noticed in both group I and group II in comparison to group III. A significant difference in lipid peroxides (LPO, P = 0.01) with highest level in patent group I (15.33 ± 0.53) followed by PCR positive latent group (14.09 ± 1.66) indicates higher lipid peroxidation in erythrocytes and oxidative stress in decreasing order when compared with infection free control horses (9.83 ± 0.97). Catalase (CAT, P = 0.01) was significantly lower in parasitological (0.82 ± 0.14) and molecular positive cases (1.27 ± 0.35) in comparison to control group (3.43 ± 0.96). The levels of superoxide dismutase (SOD, P = 0.01), reduced glutathione (GSH, P = 0.01) and ferric reducing antioxidant power (FRAP, P = 0.01) were significantly lower in parasito-molecular positive cases as compared to infection free control horses. An inverse correlation of RBC count with LPO and GSH and a direct correlation with catalase, SOD and FRAP was revealed. Overall, the observed substantial decreases in the oxidative parameters like catalase CAT, SOD, GSH and FRAP activities with remarkably elevated levels of LPO indicate high exposure of erythrocytes to oxidative damage in T.evansi infected horses.

Molecular confirmation and anthelmintic efficacy assessment against natural trichurid infections in zoo-housed non-human primates.

INTRODUCTION: This study aimed to assess Trichuris species infection and evaluate the anthelmintic efficacy of fenbendazole and ivermectin against natural trichurid infections in non-human primates (NHPs), kept at Mahendra Chaudhury (MC) Zoological Park, Chhatbir, India. MATERIALS AND METHODS: Molecular confirmation of Trichuris infection was carried out using polymerase chain reaction targeting internal transcribed spacer sequences, and anthelmintic efficacy was assessed by fecal egg count reduction test, respectively. RESULTS: A 710 base pair product confirmed Trichuris species infection in NHPs. Fenbendazole, 10 mg/kg body weight orally for 5 consecutive days and ivermectin, 100 µg/kg body weight orally for 3 alternate days proved effective and showed a maximum fecal egg reduction of 99.20% and 100% (P < .05) at day 7 post-treatment. CONCLUSIONS: This study highlighted the molecular confirmation of Trichuris species in non-human primates and its management using fenbendazole and ivermectin.

Clinical and hematobiochemical response in canine monocytic ehrlichiosis seropositive dogs of Punjab.

AIM: As in India especially, the Punjab state sero-prevalence and distribution of ehrlichiosis in relation to clinico-hematobiochemical response remains largely unexplored. Thus, this study was designed to determine the prevalence of vector (tick)-borne tropical canine pancytopenia caused by Ehrlichia canis through enzyme labeled ImmunoComb® (IC) assay in dogs from in and around Ludhiana, Punjab. Correlation of prevalence was made with various clinico-hematobiochemical parameters. MATERIALS AND METHODS: Seroprevalence study was carried out using IC® test kit (Biogal, Galed Labs). The study was conducted in 84 dogs presented to the Small Animal Clinics, Teaching Veterinary Clinical Complex, Guru Angad Dev Veterinary and Animal Sciences University, Ludhiana, Punjab. RESULTS: Out of 84 suspected dogs for ehrlichiosis, based on peripheral thin blood smear examination 12 (14.28%) cases were positive for the morulae of E. canis and 73 (86.90%) dogs were found positive to E. canis antibodies through IC® canine Ehrlichia antibody test kit, respectively. Among the different age groups 1-3 years of aged group showed highest prevalence (41.09%), followed by the 3-6 years age group (32.87%), infection levels were lower in the <1 year of age group dogs (13.69%) and more than 6 years age group dogs (12.32%). The highest prevalence was seen in Labrador retriever. This study indicates that season plays a very important role in the prevalence of ehrlichiosis. The most common findings observed were anemia, leukocytosis, neutropenia, lymphopenia, thrombocytopenia, eosinophilia followed by hyperbilirubinemia, increased levels of aspartate aminotransferase, alanine aminotransferase and alkaline phosphatase, hypoalbuminemia, hyperglobulinaemia, decrease in albumin and globulin ratio, increase in blood urea nitrogen and creatinine. CONCLUSIONS: Serological techniques like IC® are more useful for detecting chronic and subclinical infections and are ideally suited to epidemiological investigations.

Critical analysis of vector-borne infections in dogs: Babesia vogeli, Babesia gibsoni, Ehrlichia canis and Hepatozoon canis in Punjab, India.

There are few published studies on various vector borne diseases of dogs in India and most depict clinical infection in dogs, diagnosed by observation of the haemopathogens in stained blood smears. This study provides the first report regarding molecular confirmation and ancestral relationship analysis of blood smears positive cases of assorted haemopathogens in Punjab province of India. On blood smear examination, haemopathogens were observed in 124 out of 778 (15.95%, 95% CI: 13.53- 18.68) blood smears. Further polymerase chain reactions (PCR) was used on bloods smear positive cases to validate the results. Out of 778 blood samples, Babesia gibsoni was most common parasite infecting dogs (15.04%, 95% CI: 12.7-17.72), followed by Ehrlichia canis (0.39%, 95% CI: 0.0-1.13), infection of Babesia vogeli and Hepatozoon canis was same (0.26%, 95% CI: 0.0-0.9). Among various risk factors studied (age, sex, season), prevalence of infection was non-significantly higher in 1-2 year of age group (19.88%, 95% CI: 14.45-26.71), regarding sex same prevalence was recorded (15.94%), and chances of infection was highest in pre-monsoon i.e. summer (18.26%, 95% CI: 14.49-22.76). Phylogenetic analysis revealed ancestral background of Ludhiana isolates of B. vogeli, B. gibsoni, H. canis, and E. canis with the isolates of Philippines, Mongolia and Tunisia.

Histopathological and morphological studies on natural Cysticercus fasciolaris infection in liver of Wistar rats.

Descriptive morphology of multifocal hepatic cysts found in eight of forty five (17.78 %) Wistar rats sacrificed during pharmacological studies related to herbal formulations was studied. The creamish to white cysts were of varying sizes, ranging from 3-8 mm in diameter. Morphological studies of these cysts depicted the presence of metacestodes of Taenia taeniaeformis i.e. Cysticercus fasciolaris inside them. The scolex of metacestode revealed four suckers and rostellum armed with two distinct rows of characteristic pen knife shaped hooks (characteristics of taeniid cestodes). The average size of large hooks was 392.92 ± 10.12 µ and that of small hooks was 240.64 ± 14.26 µ. The average size of suckers was 304.36 ± 12.33 µ. Histopathology of hepatic tissue surrounding the cysts revealed zones of fatty change, inflammation, granulation tissue and metaplasia. However, the histopathology of stomach and small intestines didn't show any significant lesions.

Clinicopatho-Biochemical Alterations Associated with Subclinical Babesiosis in Dairy Animals.

BACKGROUND: Present investigation is based on the clinicopatho-biochemical alteration related to natural tick borne bovine babesiosis in Punjab state. METHODS: Blood samples from jugular vein of 542 bovines (cattle 466 and buffaloes 76) having history of tick infestation, fever, hemoglobinurea or anemia were collected and tested for Babesia bigemina by blood smear examination and PCR targeting 18S rRNA gene to distinguish clinically and subclinically infected groups. Further the hemato-biochemical parameters were correlated with the status of infection. RESULTS: Overall, of the 542 samples tested 16.42 % were positive by PCR while only 1.66 % by blood smear examination. The trend of molecular prevalence was found to decrease from north-eastern towards western Punjab. Analysis of the hematobiochemical alterations showed significant decrease in the levels of RBC, Hb, PCV, and MCV with significant increase in TBIL, MCH and MCHC levels. CONCLUSION: As the transmission of B. bigemina is transovarian, presence of even few infected Rhipicephalus (Boophilus) microplus ticks on a subclinically infection can be the nidus of infection for whole herd, causing severe economic losses, at the same time significantly affecting the physiology of carrier animal.

Molecular epidemiology, risk factors and hematochemical alterations induced by Theileria annulata in bovines of Punjab (India).

Bovine tropical theileriosis, caused by Theileria annulata, is one of the economically important fatal tick borne haemoprotozoan diseases of dairy animals. The aim of present investigation was to map the distribution of T. annulata in bovines of Punjab state of India in relation to various risk factors including age, sex of animals, location and management of farms. In a cross sectional study, a total of 1278 blood samples were randomly collected from twenty districts falling in five major agro-climatic zones of Punjab. All the samples were screened by blood smear examination followed by polymerase chain reaction targeting SSU rRNA gene for Theileria spp. PCR positive samples (n = 386) for Theileria spp. were then analyzed for T. annulata by amplification of Tams1 gene. Overall prevalence of T. annulata was found to be 29.26% in Punjab, with highest in western Zone (40.49%, 95% CI = 35.57-45.41) and lowest in submountain zone (18.90%, 95% CI = 13.73-24.06). The propensity of incidence of T. annulata was found to be highest in cross bred cattle (32.40%, 95% CI = 29.87-34.94), followed by indigenous cattle (19.64%, 95% CI = 10.67-28.61) and buffaloes (19.2%, 95% CI = 14.99-23.41). Between the two sexes, incidence of T. annulata was higher in female animals. Calves less than 6 months of age were found to be more prone to theileriosis.

Retrospective study of clinical and hematological aspects associated with dogs naturally infected by Hepatozoon canis in Ludhiana, Punjab, India.

OBJECTIVE: To evaluate clinical and hematological aspects of dogs naturally infected with Hepatozoon canis (H. canis) presented at the Small Animal Clinics of Guru Angad Dev Veterinary and Animal Sciences University, Ludhiana. METHODS: Blood films of 34 naturally infected dogs were examined for haematological alterations and parasitaemia. Signalment and clinical signs were recorded from the animals. Clinical histories were filled out during the consultation. RESULTS: Of the 34 positive dogs by Giemsa stained peripheral blood films, 88.23% presented parasitaemia by H. canis only, while 11.77% had the combination of H. canis, Babesia sp. and Ehrlichia sp. Young male dogs less than one-year-old, of non-descript breed, were the most commonly affected. And 26.47% were presented with anorexia/inappetence as the only clinical symptom. Other clinical symptoms were mild to moderate fever, pale mucosae and lethargy; a few were also showing the signs of vomiting and diarrhoea. Haematological alterations showed mainly normochromic-normocytic anaemia, leukocytosis and neutrophilia. CONCLUSIONS: The findings of this study substantiate that H. canis caused clinical and haematological alterations of the varied intensity in dogs, even with low parasitaemia, should be taken into consideration.

Prevalence and haemato-biochemical profile of Anaplasma marginale infection in dairy animals of Punjab (India).

OBJECTIVE: To do the systematic comparison of prevalence of anaplasmosis by PCR and Giemsa stained thin blood smear (GSTBS) based parasitological assays in dairy cattle of Punjab, which has not been reported yet. To analyse the haematobiochemical alterations in infected animals to arrive at the conclusion regarding the pathogenicity induced by Anaplasma marginale (A. marginale) in latent and patent infection. METHODS: Study was conducted on 320 animals (236 cows, 62 calves and 22 buffaloes) of Punjab, India. PCR on genome of A. marginale was performed by targeting msp1 ß gene using specific primers BAP-2/AL34S, amplifies products of size 407 bp. Questionnaires based data on the characteristics of the infected animals and management strategies of the farm were collected and correlated. RESULTS: Higher prevalence and more significant association was observed in the PCR based molecular diagnosis (P=0.00012) as compared to that in GSTBS (P=0.028 8) based diagnosis with various regions under study. With respect to the regions, highest prevalence was recorded in Ferozepur by PCR based diagnosis, while that in Jalandhar by GSTBS examination. Similar marked significant association of the PCR based diagnosis with the age of the animals under study (P=0.00013) was observed elucidating no inverse age resistance to A. marginale in cow calves. Haematobiochemical profile of infected animals revealed marked anemia, liver dysfunction and increase globulin concentrate indicating rise in immunoglobulin level to counteract infection. CONCLUSIONS: PCR is far more sensitive in detecting the disease even in latent infection which may act as nidus for spread of anaplasmosis to susceptible animals in endemic areas. Severity of anaemia and liver dysfunction were comparable both in patent as well as latent infection indicating pathogenicity of both.

Vaccines against Toxoplasma gondii: status, challenges and future directions.

Toxoplasma gondii is a ubiquitous protozoan parasite that can infect a wide range of animals including humans. This single known species in the genus Toxoplasma is considered as one of the most successful eukaryotic pathogens which is of major medical and veterinary importance. Effective vaccines may contribute toward preventing and controlling the spread of toxoplasmosis. The present communication addresses the current status of development of vaccines against T. gondii. Further discussion is made on the difficulties along with challenges, such as vaccine construct, mode of vaccine administration and standardization of immunization evaluation. Finally suggestions are made on possible directions for future research on the development of vaccines against T. gondii.