Sarcocystosis is an emerging zoonotic disease caused by apicomplexan parasites of the genus Sarcocystis, which can infect a wide range of hosts, including humans, animals, reptiles, and birds. Humans may serve as either definitive or intermediate hosts, depending on Sarcocystis species. Consumption of undercooked beef containing mature sarcocysts of Sarcocystis hominis or Sarcocystis heydorni, or of raw or partially cooked pork containing Sarcocystis suihominis zoitocysts, may lead to intestinal sarcocystosis in humans. In contrast, muscular sarcocystosis follows ingestion of food or water contaminated with zoonotic Sarcocystis spp. sporocysts, as evidenced by infection outbreaks by Sarcocystis nesbitti among travelers returning from exotic destinations in Malaysia. Further, Sarcocystis fayeri, which cycles between horses and dogs, has been implicated in cases of food poisoning in humans following consumption of raw horse meat. Clinical features range from asymptomatic cases to acute symptoms, such as muscle pain, fever, and, in severe cases, neurological complications. Diagnosis relies on clinical presentation supported by microscopy, histopathology, serology, and advanced molecular techniques. Therapeutic options are limited; antiprotozoal agents including sulfadiazine, pyrimethamine, decoquinate, clindamycin, and triazine derivatives have shown high efficacies. Preventive measures focus on hygienic meat handling, thorough cooking, and restricting carrion scavenging by stray carnivores. Increasing human-wildlife interactions elevate the risk of cross-species transmission of novel Sarcocystis species. Adopting a One Health approach is fundamental for understanding the epidemiology and transmission dynamics of zoonotic sarcocystosis and reducing future outbreaks through effective diagnostics, therapeutics, and preventive interventions.

Poultry has protozoa that are classified into multiple taxonomic groupings. In poultry, two types of parasites are significant: the coccidia and the mastiogophora (flagellates). Some parasites, which cause coccidiosis, have short, direct life cycles and are therefore preferred, while other parasites that involve intermediate hosts typically do not pose a threat to commercial poultry. A significant exception is blackhead disease (histomoniasis), which has a complex life cycle involving intermediate hosts, but relies on chickens as reservoir hosts and spreads easily among turkeys within a flock. Most coccidia found in poultry belong to the genus Eimeria, however, there are also some species of Isospora and Cryptosporidium. The most well-known are the Eimeria, of which seven significant species have been identified in chickens and several more in turkeys. Anywhere chickens are raised, whether in huge commercial operations or tiny backyard flocks, parasites are an issue that can result in severe financial losses. This chapter will provide a quick overview of the main poultry protozoan parasitic species, along with some pathophysiology.

The use of diagnostic methods for the diagnosis of parasitic diseases in poultry has been almost constant over the past few decades. Since the introduction of PCR, few major advances have been adopted in clinical diagnostic tests. Many diagnostic tests that form the backbone of the “modern” microbiology laboratories rely on very old and labour-intensive technologies such as microscopy for the diagnosis of parasites including helminths, protozoans, arthropods, and haemoprotozoans. Urgent needs include more rapid tests without compromising the sensitivity, value-added tests, and point-of-care tests for both high- and low-resource settings. In recent years, research has been focused on alternative methods to improve the diagnosis of parasitic diseases. These include molecular technique-based approaches, immunoassays and proteomics using mass spectrometry platforms technology. This chapter discusses the progress of several approaches in parasite diagnosis and some of their silent characteristics.

Trypanosoma evansi, a hemoflagellate protozoan, leads to wasting disease, surra in livestock animals causing huge economic losses. Currently, the preferred assay for surra diagnosis is whole cell lysate (WCL) based ELISA, which requires the use of rodents for WCL preparation. To avoid use of laboratory animals, we used recombinant DNA technology to express T. evansi invariable surface glycoprotein (ISG) in E. coli. The potential of recombinant ISG65 (rISG65) as a diagnostic antigen was investigated in immunoblot and indirect ELISA using experimentally infected equine serum samples from 0 to 84 days post infection. The results indicated that rISG65 reacted with horse T. evansi positive serum giving two bands of approximately 48 kDa and 96 kDa. T. evansi-specific antibodies were detected as early as 10 and 14 days post infection using immunoblot and indirect ELISA, respectively using rISG65 antigen. No cross-reactivity was observed in ELISA and immunoblot with different serum samples of equines positive for Equine herpesvirus 1, Burkholderia mallei, and Theileria equi infections. Several immunoreactive regions were observed between 30 and 100 kDa in T. evansi isolate of horse origin indicating the existence of multiple copies of ISG protein in a single trypanosome. The recombinant ISG has proven to be good candidate antigen to be used in ELISA for serodiagnosis of T. evansi infection in different animals.

Serodiagnosis of surra, caused by Trypanosoma evansi, is still based on native antigens purified from bloodstream form of T. evansi grown in rodents. In order to investigate prospective diagnostic possibilities as an alternative for native antigens, we cloned, expressed 26 kDa calflagin protein containing 218 amino acids from T. evansi (Indian Strain) in Escherichia coli. The potential of recombinant calflagin (rCLF) protein as diagnostic antigen was evaluated in immunoblot and indirect ELISA using experimentally infected equine serum samples from 0 to 84 days post infection. The antibodies against T. evansi were detected with rCLF antigen in serum samples of experimentally infected equines as early as 10 days and 14 days post infection, using immunoblot and ELISA respectively. No cross-reactivity was observed with rCLF antigen in ELISA with different serum samples of equines positive for Equine herpesvirus 1, Burkholderia mallei, and Theileria equi infections. Several immunoreactive regions ranging from 10 to 28 kDa were detected using distinct T. evansi isolates (pony, cattle, donkey and camel origin) indicating presence of multiple calflagin family members in a single trypanosome. Indirect immunofluorescence antibody test with anti-CLF rabbit hyperimmune serum showed localisation of native immunogenic protein near attachment of flagellum. The rCLF protein was found to be a potential diagnostic candidate for distinguishing T. evansi positive and negative equine serum sample, suggesting that it could be used for serological surveys in animals for surra. In addition, it could be used with other potential diagnostic candidates to improve the diagnostic efficiency.

Background: Procrastination is considered problematic when it affects normal day-to-day functioning. It can serve as a deterrent among undergraduate dental students both for their academic and career growth. Behaviour modifications are required to change this negative behaviour and Sense of Coherence is one such approach. Aim: To assess the relationship between Sense of Coherence and Procrastination among a group of undergraduate dental students in Chennai city. Material and Methods: A cross sectional questionnaire study was conducted among 358 undergraduate dental students. Thirteen item Sense of Coherence scale and Lay’s Procrastination scale were used to collect data about Sense of Coherence and Procrastination among undergraduate dental students respectively, using google forms. One-way ANOVA and Independent T test were used to test for difference in mean scores between groups based on year of study and gender, respectively. Pearsons correlation coefficient followed by Simple Linear Regression was performed to test for association between Sense of Coherence and Procrastination. Results: The overall mean Sense of Coherence score of the study participants was 49.52 ± 8.13. The overall mean Procrastination score of the study participants was 56.58 ± 10.02. Pearson’s Correlation Coefficient revealed a negative correlation (r = -.255) between Sense of Coherence and Procrastination (p = .000) indicating that with an increase in Sense of Coherence, Procrastination decreases. In tandem, the third year students with lowest Sense of Coherence score showed highest procrastination. Conclusion: Procrastination is negatively associated with Sense of Coherence and hence behaviour modification interventions based on Sense of Coherence are required, to be customized and incorporated in dental curriculum to reduce procrastination among undergraduate dental students.

The development of the tumorigenesis and angiogenesis through proteolytic cleavage of extracellular matrix protein and basement membranes is promoted by Matrix metelloproteinases-7 (MMP-7). Consequently, MMP-7 is presumed as potential target for mammary cancer immunotherapy. However, MMP-7 is an endogenous tumor associated antigen (TAA); therefore, immunization is challenging. In current study, a potent anti-tumor immune response has been elicited through recombinant bivalent plasmid pVIVO2.IL18.cMMP7 which subside the highly metastatic 4 T1 cell line induced mammary tumors and efficiently negate the existing challenge of using MMP-7 as immunotherapeutic target. Balb/c mice were immunized with canine MMP-7 (cMMP-7) using interleukine-18 (IL-18), as an immunoadjuvant, to explore the potential of the combination regarding elicitation of a potent anti-tumor immune response. Mice vaccinated with pVIVO2.IL18.cMMP7 DNA plasmid reduced the tumor growth significantly along with augmentation of the immune response to fight against tumor antigen as depicted by substantial enrichment of CD4+ and CD8+ population in splenocytes, infiltration of immune system cells in tumor tissue and enhanced survival time of mice. Further, splenocyte supernatant examination of the cytokines revealed that Th1 cytokines (IFN-γ and IL-2) were remarkably up-regulated demonstrating the stimulation of cell-mediated immune response. Thus the current observations vividly portray that administration of xenogeneic MMP-7 DNA vaccine bypasses the tolerance barrier.

Trypanosoma evansi, an extracellular haemoflagellate, has a wide range of hosts receptive and susceptible to infection, in which it revealed highly inconsistent clinical effects. Drugs used for the treatment of trypanosomosis have been utilized for more than five decades and have several problems like local and systemic toxicity. In the present investigation, imatinib and sorafenib were selected as drugs as they are reported to have the potential to cause reactive oxygen species (ROS)–mediated effect in cancer cells. Both have also been reported to have potential against T. brucei, T. cruzi and Leishmania donovani. To date, imatinib and sorafenib have not evaluated for their growth inhibitory effect against T. evansi. Imatinib and sorafenib showed significant (p < 0.001) inhibition on parasite growth and multiplication with IC50 (50% inhibitory concentration) values 6.12 μM and 0.33 μM respectively against T. evansi. Both the drug molecules demonstrated for the generation of ROS in T. evansi and were found up to 65% increased level of ROS as compared with negative control in the axenic culture system. Furthermore, different concentrations of imatinib and sorafenib were found non-toxic on horse peripheral blood mononuclear cells and Vero cell lines. Also, in conclusion, our results demonstrated that imatinib- and sorafenib-induced generation of ROS contributed inhibitory effect on the growth of Trypanosoma evansi in an axenic culture system.

Advent of quantitative polymerase chain reaction and its variants have enabled identification and quantification of seven known Eimeria species of poultry in biological samples. Attempts were made in the present study to identify and quantify three important pathogenic Eimeria species responsible for intestinal coccidiosis in domestic farmed chicken, E. necatrix, E. acervulina and E. maxima in droppings collected from thirty one poultry farms of North Indian states of Haryana, Punjab, Uttar Pradesh and Uttarakhand. The study included broiler, layer and backyard rearing units. Overall occurrence of E. necatrix, E. maxima and E. acervulina was 64.5%. E. necatrix was detected in 55% (11/20) broiler farms, 66.7% (4/6) layer farms and 100% (5/5) backyard rearing units studied. Thus, occurrence of E. necatrix was detected in 64.5% (20/31) farms studied. E. maxima and E. acervulina were detected in droppings of 65% (13/20) broiler farms, 66.7% (4/6) layer farms and 60% (3/5) back yard rearing units. Genome counts of each Eimeria species revealed maximum parasite load of E. necatrix followed by E. acervulina in broiler farms and least in layer farms. The mean parasite load (genome) copies for these parasite species were intermediate for backyard units while E. maxima had the lowest number of genome copies in droppings. Mean E. maxima counts were highest in boiler farms, while it was similar for layer and back yard units. However, statistically no significant differences were observed for parasite load existing either between the broiler, layer or back yard units or between the genome counts of E. necatrix, E. acervulina or E. maxima.

Six Trypanosoma evansi isolates were collected from ponies (PH1 and PK6), camel (CB2), donkeys (DJ3 and DH4) and cattle (CK5) from different States of Northern India (Haryana, Rajasthan, Uttar Pradesh and Gujarat) for molecular characterization based on 18S rRNA gene. The 18S rRNA gene (2251 bp) of different isolates was amplified, cloned and custom sequenced separately. Based on sequence and phylogenetic analysis of all six isolates, collected from different hosts as well as geographical areas, were having high identity among Indian T. evansi strains (99.7%) and with other strains of T. evansi (99.2%) distributed worldwide. There is less genetic diversity among different salivarian strains of T. evansi except few nucleotide changes at significant locations in one Indian isolate of camel origin (CB2). All Indian T. evansi isolates were grouped in salivarian clade with high bootstrap values and remained far away from stercorarian clade having 88-90% nucleotide identity. The study will be helpful in understanding the evolutionary relationship, molecular epidemiology and variation in disease pathogenesis among different T. evansi strains. Further, more studies are required on large number of isolates collected from diverse host and geographical areas to reaffirm the present finding.

This study is based on how to engage the employee in their work place and to improve for their further career development. In this study primary data and secondary data were collected. Standard questionnaire and personnel interviews were used to collect primary data from the individuals. Secondary data were collected from company broachers, magazines, websites

The present study was undertaken to identify and purify the immunodominant fractions from the excretory secretory (ES) antigen of Paramphistomum epiclitum, a predominant amphistome species infecting ruminants in India. ES antigen was prepared and characterized using SDS-PAGE and Western blot analysis. Major polypeptides of molecular weight 11, 22, 28, 31, 33, 39, 52, 59, 63 and 72 kDa were visualized in SDS-PAGE. Polypeptides (9) of 11, 14, 16, 22, 31, 33, 39, 63 and 72 kDa showed immunoreactivity in Western blot analysis. The whole ES antigen of P. epiclitum was initially concentrated using PEG-8000 followed by spin-X UF concentrator with 10 kDa cutoff range and subsequently fractionated by size exclusion chromatography using Sephadex G-25. Cross reactivity of the P. epiclitum ES antigen was studied with positive sera of F. gigantica and H. contortus. Based on the cross reactivity profile, the low molecular weight antigenic fraction with 11 kDa polypeptide was selected for further use in indirect-ELISA. Bovine serum samples (258) were tested with optimized ELISA. Sensitivity of the ELISA was calculated as 75.0%, while the specificity was 85.0%. The percent positive and negative predictive values for the test were 70.78 and 87.57%, respectively.

Parasitic diseases account for important health hazard in man and animal in tropical countries like India. Ectoparasites infestation causes a serious loss in health and economy every year in India. They can cause annoyance, irritation, skin infection, anaemia, tick fever as well as act as a vector for various devastating diseases. Thus, ectoparasites control is a matter of great concern. Various chemical acaricides have been prescribed since last 50 years. But, their residual effect, adverse side effect, and resistance are a matter of concern now days. Hence, biological control of ectoparasites gains prime importance in many parasite control program. The present review critically analyzes the different methods for controlling the ectoparasites with special emphasis on the newer approach of the biological, immunological, genetic and pheromone method.

Matrix metalloproteinases-7 (MMP-7) which is expressed in a wide variety of malignant cells has been seen tobe extensively up-regulated in mammary carcinomas. MMP-7 can promote cancer invasion and angiogenesis through proteolytic cleavage of extracellular matrix and basement membrane proteins. This property of MMP-7 makes it a promising target in the context of immunotherapy. Further, to enhance DNA-based immunization, a cytokine gene can be employed as an adjuvant. Interleukin-18 (IL-18) is a Th1-type cytokine that has been demonstrated as a potential biological adjuvant in murine tumour models. The present study was undertaken to clone murine MMP-7 (mMMP-7) and IL-18 genes in pVIVO2.mcs eukaryotic expression vector and to characterize their expression by immunofluorescence and Western blotting. This double gene construct now may be used as a potential xenogeneic DNA vaccine against canine tumour model.

A wealth of genomic and proteomic information on microorganisms and parasites, together with recent advances in adjuvant and delivery systems, is being harnessed to develop nanovaccines against infectious and parasitic diseases. The use of nanoparticles in vaccine formulations allows not only improved antigen stability and immunogenicity but also targeted delivery and slow release. However, so far, nanoparticles have not proved capable of surmounting most of the barriers like toxicity issues, clearance from biological system, DNA instability and differences in expression systems. Nevertheless, advances in nanoparticle engineering and understanding of nanoparticle characteristics, is creating new opportunities for the development of nanovaccines as antigen delivery system. This Review focuses on recent progress in development of nanoparticle based antigen delivery vehicles, their use in different diseases, major bottlenecks and challenges to realizing the potential of nanoparticles.

Most of the bacterial diseases are sporadic rather than epidemic in equines as far as the affections of nervous system are concerned. Bacteria of genus Streptococcus often leads to meningitis or abscess in brain. In one of the study Staphylococcus spp. was found to be involved as a major cause of encephalitis [1] while some researchers investigated E. coli, Streptococcus spp., Staphylococcus spp., Listeria and Actinobacillus as major contributors for encephalitis and encephalomyelitis in horses [2]. Streptococcus spp., Staphylococcus aureus and Gramnegative enteric bacteria are most frequently isolated causes of encephalitis in foals [3]. Most of the bacterial affections of brain in foals are because of lack of humoral immunity and increased permeability of BBB [4]. However, Streptococcus equi subspp equi, Zoopedemicus, Streptococcus suis, Klebsiella pneumoniae, Pasteurella caballi, Actinobacillus equuli, Actinomyces spp. and E.coli [5-7] are identified as major bacterial pathogens associated with encephalitis in adult horses. In many of the cases causes of meningitis are not clear [8]. The lesions in brain can be associated with some surgical interventions or external injuries as well. The meningitis and encephalitis may be found to occur by skull fracture, pituitary abscess (as fenestrated BBB) and extension of infection from adjacent organs like guttural pouch or retropharyngeal lymph nodes etc. [8]. Meningitis is mostly secondary to brain abscess [8, 9]. Diagnosis of bacterial meningitis can be made on the basis of clinical examination, where neutrophilia is most significant feature along with increase fibrinogen level in blood [10] and blood culture also indicate positive results if any infection is present [11]. Abstract The nervous system affections are one of the major reason for death in equines. These affections can involve a huge spectrum of etiologies namely viral, bacterial, parasitic, fungal, algal, plant toxins, mycotoxins etc. Botulinum and tetanus are among most commonly known deadly conditions affecting nervous system in equines. Botulinum (BoNT) and Tetanospasmin are the most potent known neurotoxins respectively which are majorly involved in nervous system damage. However certain parasitic diseases like EPM and Toxoplasma are also involved in many encephalitic conditions. Cryptococcus is among one of the most important known fungus causing encephalitis in equids. Often certain plants like Pteridium aquilinum and Locoweed etc. ingested by equines lead to lesions in brain and the toxins present in these weeds are having different specificity for different parts of brain e.g. Cerebrocortical necrosis (3 rd , 5 th & 6 th layer of cerebrum) by Pteridium aquilinum and injury to Purkinje cells and cerebral cortex by Locoweed. In this article we are mainly focusing on certain bacterial, parasitic, fungal, algal and plant toxicity induced conditions causing encephalitis in equines as these conditions are often overlooked.

Animal trypanosomosis (surra), caused by Trypanosoma evansi, is one of the most important diseases in livestock and wild animals in India. The disease is prevalent across all agro-climatic regions of India, and has a considerable impact on the country's livestock economy through direct and indirect impact on livestock productivity. In the present study, the economic losses on livestock productivity were assessed resulting from surra in India, considering all possible direct and indirect losses in major six livestock species viz., cattle, buffalo, goat, equine, camel and pig. The contemplative risk and retrospective analyses were performed using various official records and scientific literature complemented with expert data for evaluation of impact of surra on livestock productivity in India. Most of the information were derived using the secondary data published in scientific journals, and the official data reported by Basic Animal Husbandry and Fisheries Statistics (BAHS, 2014), the Government of India and other scientific reports. To address the variability and uncertainty, probability distributions for many input values were used in the present study and sensitivity statistical analyses were conducted using a simulation model. In the current analysis, all prices were assumed as deterministic. Based on present study, a total annual loss (direct and indirect) caused by surra was estimated to US $ 671.1 million or Indian Rupee (INR) 44,740 million (US $ 344–US $ 1209 million or INR 22951.88–80,752.35 million at 95% confidence interval), at present valuation. The mortality losses were estimated to 15.67% of the total loss. Among morbidity losses, the reduction in milk yield and reproductive losses components were 36.46% and 25.72% of total loss, respectively. Other parameters like reduction in growth (9.83%), reduction in draught power (7.95%) and additional opportunity cost (2.93%) also yielded considerable loss. The results highlighted the urgent need for early diagnosis and control strategies for surra in livestock species to reduce the productivity losses in the country's livestock sector.

Eimeria tenella, the causative agent of caecal coccidiosis, is a pathogenic gut dwelling protozoan which can cause severe morbidity and mortality in farmed chickens. Immune mapped protein-1 (IMP-1) has been identified as an anticoccidial vaccine candidate; in the present study allelic polymorphism was assessed across the IMP-1 coding sequence in E. tenella isolates from four countries and compared with the UK reference Houghton strain. Nucleotide diversity was low, limited to expansion/contraction of a CAG triplet repeat and five substitutions, three of which were non-synonymous. The EtIMP-1 coding sequence from a cloned Indian E. tenella isolate was expressed in E. coli and purified as a His-tagged thioredoxin fusion protein. An in-vivo vaccination and challenge trial was conducted to test the vaccine potential of recombinant EtIMP-1 (rEtIMP-1) and to compare post-vaccination immune responses of chickens to those stimulated by live oocyst infection. Following challenge, parasite replication measured using quantitative PCR was significantly reduced in chickens that had been vaccinated with rEtIMP-1 (rIC group; 67% reduction compared to UC or unimmunised controls; 79% reduction compared to rTC group or recombinant thioredoxin mock-immunised controls, p<0.05), or the birds vaccinated by infection with oocysts (OC group, 90% compared to unimmunised controls). Chickens vaccinated with oocysts (OC) had significantly higher levels of interferon gamma in their serum post-challenge, compared to rEtIMP-1 vaccinated birds (rIC). Conversely rEtIMP-1 (rIC) vaccinated birds had significantly higher antigen specific serum IgY responses, correlating with higher serum IL-4 (both p<0.05).

Indirect ELISA and dot-ELISA using recombinant BgP12 (rBgP12) were developed for the diagnosis of Babesia gibsoni infected dogs. The complete open reading frame of BgP12 gene (378 bp) was cloned in pET-32a(+) expression vector and expressed in Escherichia coli as a soluble thioredoxin (Trx) fusion protein. The purified rBgP12 was used for production of anti-rBgP12 rabbit serum, which recognized a native 12-kDa protein in B. gibsoni infected erythrocyte by Western blot analysis. To evaluate the potential of rBgP12 for the serodiagnosis of B. gibsoni , a panel of serum/plasma samples from dogs infected with B. gibsoni ( n = 13), uninfected sera ( n = 13) and sera from dogs infected with other haemoparasites viz. , Babesia canis vogeli ( n = 3), Ehrlichia canis ( n = 3), Hepatozoon canis ( n = 1) and Dirofilaria immitis ( n = 1) were used in ELISA formats. In addition, the performance of rBgP12 based indirect ELISA and dot-ELISA were evaluated using 75 serum/plasma samples collected from suspected dogs, in respect to the nested PCR as reference test. The diagnostic sensitivities of indirect ELISA and dot-ELISA were 94.59% and 89.18%, respectively, while their specificities were 84.21% and 81.57%, respectively. Moreover, both the assays using rBgP12 showed no cross reaction with sera from dogs infected with other common haemoparasites indicating their high specificity. High kappa values of indirect ELISA and dot-ELISA indicated the potentials of these assays with substantial agreement at 95% confidence level. It is concluded that indirect ELISA and dot ELISA using rBgP12 might be used in large scale epidemiological surveys and clinical diagnosis of B. gibsoni infection in dogs.