Neuropharmacological Effects of Chassalia curviflora (Rubiaceae) Leaves in Swiss Albino Mice Model.

The current study aimed to investigate the neuropharmacological properties of ethanol, acetone, and ethyl acetate leaf extracts of Chassalia curviflora (C. curviflora) in mouse models. The neuropharmacological properties of this plant were studied on Swiss albino mice at dosages of 50, 100, and 200 mg/kg body weight in thiopental sodium-induced sleeping time test, and at dosages of 100 and 200 mg/kg body weight in other tests. The extracts caused a marked reduction in the initiation and sleep length (P<0.05) in studies on thiopental sodium-induced sleeping time at dosages of 100 and 200 mg/kg and a significant decrease (P<0.05) was found in terms of unconstrained locomotor and explorative activities in both hole crossing and open field tests at dosages of 100 and 200 mg/kg. Furthermore, the extracts increased sleeping time with a dosage-dependent onset of action. The hole-board test extracts also reduced the number of head dips at dosages of 100 and 200 mg/kg (P<0.05). It was found in this study that C. curviflora had the best neuropharmacological properties at a dosage of 200 ml/kg. Our findings also showed that all of the extracts from C. curviflora were experimentally active in an in vivo model. The study results suggested that the leaves had strong anti-depressant and hypnotic CNS properties that might be exploited for neuropharmacological adjuvant therapy in conventional medicine. However, pharmacological studies are warranted to explore the active substances and the mode of action.

Relooking the monkeypox virus during this present outbreak: epidemiology to therapeutics and vaccines.

OBJECTIVE: The recent monkeypox disease outbreak is another significant threat during the ongoing COVID-19 pandemic. This viral disease is zoonotic and contagious. The viral disease outbreak is considered the substantial infection possessed by the Orthopoxvirus family species after the smallpox virus' obliteration, a representative of the same family. It has potentially threatened the Republic of Congo's regions and certain African subcontinent zones. Although repeated outbreaks have been reported in several parts of the world, as conferred from the epidemiological data, very little is explored about the disease landscape. Thus, here we have reviewed the current status of the monkeypox virus along with therapeutic options available to humanity. MATERIALS AND METHODS: We have accessed and reviewed the available literature on the monkeypox virus to highlight its epidemiology, pathogenicity, virulence, and therapeutic options available. For the review, we have searched different literature and database such as PubMed, PubMed Central, Google Scholar, Web of Science, Scopus, etc., using different keywords such as "monkeypox", "Orthopox", "smallpox", "recent monkeypox outbreak", "therapeutic strategies", "monkeypox vaccines", etc. This review has included most of the significant references from 1983 to 2022. RESULTS: It has been reported that the monkeypox virus shows a remarkable similarity with smallpox during the ongoing outbreak. Sometimes, it creates considerable confusion due to misdiagnosis and similarity with smallpox. The misdiagnosis of the disease should be immediately corrected by rendering some cutting-edge techniques especially intended to isolate the monkeypox virus. The pathophysiology and the histopathological data imply the immediate need to design effective therapeutics to confer resistance against the monkeypox virus. Most importantly, the potential implications of the disease are not given importance due to the lack of awareness programs. Moreover, specific evolutionary evidence is crucial for designing effective therapeutic strategies that confer high resistance, particularly against this species. CONCLUSIONS: The review focuses on a brief overview of the recent monkeypox virus outbreak, infection biology, epidemiology, transmission, clinical symptoms, and therapeutic aspects. Such an attempt will support researchers, policymakers, and healthcare professionals for better treatment and containment of the infection caused by the monkeypox virus.

Grading of Anatomopathological Disparity in the Cases of Invasive Pulmonary Aspergillosis in wild avian species as recorded in Pigeons (Columba livia), Peafowls (Pavo cristatus), and Griffon Vultures (Gyps fulvus).

Aspergillosis which is caused by Aspergillus fumigatus, a fungal pathogen, can vary from a localized infection to severe life-threatening invasive or disseminated systemic diseases in birds. The present study aimed to evaluate and grade the anatomopathological disparity in the cases of invasive pulmonary aspergillosis (IPA) in Columba livia (pigeons), Pavo cristatus (peafowls), and Gyps fulvus (Griffon vultures). Necropsy gross lesions varied from mere congestion of lungs in P. cristatus, congestion and large necrotizing masses surrounded by a zone of hyperemia (10 mm dia) in lungs of C. livia, and typically disseminated granuloma in the lungs, air sacs, and organs of other serous membranes in G. fulvus. Histopathology varied from extensive parenchymal necrosis amidst exuberant fungal invasion in P. cristatus, multifocal to focally extensive tissue necrosis with colonies of fungal hyphae surrounded by heterophils and lymphocytes in C. livia, as well as typical mycotic granuloma embedded in the lungs, air sacs, and thoracoabdominal serous membranes with angio-invasion in G. fulvus. Based on gross and histopathological findings, we diagnosed the cases as Acute Invasive Pulmonary Aspergillosis (AIPA) in peafowls and pigeons, as well as Chronic Invasive Pulmonary Aspergillosis (CIPA) in Griffon vultures. There is a paucity of case reports on aspergillosis in wild avian species, and this report strived to document the cases of IPA in peafowls, pigeons, and vultures. This is the first report of its kind which evaluated anatomopathological disparity of IPA in pigeons, peafowls, and vultures with a proposed anatomopathological grading system which would help to understand and investigate the nature of aspergillosis in different avian hosts.

Comparative pathology, molecular pathogenicity, immunological features, and genetic characterization of three highly pathogenic human coronaviruses (MERS-CoV, SARS-CoV, and SARS-CoV-2).

The last two decades have witnessed the emergence of three deadly coronaviruses (CoVs) in humans: severe acute respiratory syndrome coronavirus (SARS-CoV), Middle East respiratory syndrome coronavirus (MERS-CoV), and severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). There are still no reliable and efficient therapeutics to manage the devastating consequences of these CoVs. Of these, SARS-CoV-2, the cause of the currently ongoing coronavirus disease 2019 (COVID-19) pandemic, has posed great global health concerns. The COVID-19 pandemic has resulted in an unprecedented crisis with devastating socio-economic and health impacts worldwide. This highlights the fact that CoVs continue to evolve and have the genetic flexibility to become highly pathogenic in humans and other mammals. SARS-CoV-2 carries a high genetic homology to the previously identified CoV (SARS-CoV), and the immunological and pathogenic characteristics of SARS-CoV-2, SARS-CoV, and MERS contain key similarities and differences that can guide therapy and management. This review presents salient and updated information on comparative pathology, molecular pathogenicity, immunological features, and genetic characterization of SARS-CoV, MERS-CoV, and SARS-CoV-2; this can help in the design of more effective vaccines and therapeutics for countering these pathogenic CoVs.

Modulation of host epigenome by coronavirus infections and developing treatment modalities for COVID-19 beyond genetics.

The recent Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2) outbreak has resulted in coronavirus disease 2019 (COVID-19) pandemic worldwide, affecting millions of lives. Although vaccines are presently made available, and vaccination drive is in progress to immunize a larger population; still the risk of SARS-CoV-2 infection and related mortality is persistent amid threats of the third wave of the ongoing pandemic. In the scenario of unavailability of robust and efficient treatment modalities, it becomes essential to understand the mechanism of action of the virus and deeply study the molecular mechanisms (both at the virus level and the host level) underlying the infection processes. Recent studies have shown that coronaviruses (CoVs) cause-specific epigenetic changes in the host cells to create a conducive microenvironment for replicating, assembling, and spreading. Epigenetic mechanisms can contribute to various aspects of the SARS-CoV-2 multiplication cycle, like expressing cytokine genes, viral receptor ACE2, and implicating different histone modifications. For SARS-CoV-2 infection, viral proteins are physically associated with various host proteins resulting in numerous interactions between epigenetic enzymes (i.e., histone deacetylases, bromodomain-containing proteins). The involvement of epigenetic mechanisms in the virus life cycle and the host immune responses to control infection result in epigenetic factors recognized as emerging prognostic COVID-19 biomarkers and epigenetic modulators as robust therapeutic targets to curb COVID-19. Therefore, this narrative review aimed to summarize and discuss the various epigenetic mechanisms that control gene expression and how these mechanisms are altered in the host cells during coronavirus infection. We also discuss the opportunities to exploit these epigenetic changes as therapeutic targets for SARS-CoV-2 infection. Epigenetic alterations and regulation play a pivotal role at various levels of coronavirus infection: entry, replication/transcription, and the process of maturation of viral proteins. Coronaviruses modulate the host epigenome to escape the host immune mechanisms. Therefore, host epigenetic alterations induced by CoVs can be considered to develop targeted therapies for COVID-19.

Variants of SARS-CoV-2, their effects on infection, transmission and neutralization by vaccine-induced antibodies.

OBJECTIVE: The current study reviewed Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2) variants for their effects on infection, transmission and neutralization by vaccine-induced antibodies. MATERIALS AND METHODS: The research articles for the current study were searched over PubMed, Google Scholar, EMBASE and Web of Science online databases. The keywords used were: (("SARS-CoV-2" OR "COVID-19") AND ("mutation" OR "variant") AND ("death" OR "hospitalization" OR "infection" OR "transmission") AND ("antibody" OR "neutralize" OR "vaccine")). A total of 333 research articles were retrieved through online-database search. These articles were further scrutinized for their relevancy. Additionally, searches were performed to find the latest relevant information over Google search engine and relevant news browsers. Finally, around 35 germane articles were considered for scripting the current report. RESULTS: The mutations have changed amino acids at key positions in spike protein viz. S477N, E484K, Q677H, E484Q, L452R, K417T, K417N and N501Y. These mutations are relevant for different characteristics and are present in newly evolved strains of SARS-CoV-2 like E484K in B.1.526, B.1.525, P.2, B.1.1.7, P.1 and B.1.351. Mutations have increased the immune escape potential leading to 3.5-6.5-folds decrease in neutralization of antibodies (Pfizer and Moderna vaccines). The variant, B.1.617 circulating in India and many other countries (double variant) having E484Q and L452R mutations, has raised the infection rate and decreased the neutralization capacity of the vaccine-induced antibodies. Deadly K417N+E484K+N501Y triplet mutations found in B.1.351 and P.1 have increased the transmission ability of these strains by 50% leading to greater COVID-19 hospitalization, ICU admissions and deaths. CONCLUSIONS: The new SARS-CoV-2 variants have compromised the neutralization potential of the currently used vaccines, but still, they have considerable efficacy to reduce infection and mortality. GRAPHICAL ABSTRACT: https://www.europeanreview.org/wp/wp-content/uploads/Graphical_Abstract.jpg.

Percutaneous transplantation of allogenic bone marrow-derived mesenchymal stem cells for the management of paraplegia secondary to Hansen type I intervertebral disc herniation in a Beagle dog.

BACKGROUND: Intervertebral disc herniation (IVDH) is one of the common causes of spinal cord injury (SCI) in dogs. It is commonly treated by performing surgical decompression that involves the removal of the extruded disc material. However, the recovery rates after surgical interventions are variable and many times unsatisfactory. This report aims to document a case of paraplegia associated with IVDH in a Beagle dog and its therapeutic management using allogenic bone marrow-derived mesenchymal stem cells (aBM-MSCs). CASE DESCRIPTION: The dog was presented with paraplegia that was initiated three weeks back. Based on the findings of computed tomography (CT), the condition was diagnosed as Hansen type I IVDH at T12-T13 intervertebral space. FINDINGS/TREATMENT AND OUTCOME: Neurological examination was performed to grade the neurological deficit. The isolation, culture, and characterization of aBM-MSCs were done as per the standard protocol. The prepared cell suspension of aBM-MSCs was percutaneously transplanted to the spinal cord parenchyma at the site of injury. A total of four doses of 1 × 106 cells were given at an interval of 15 days along with methylcobalamin and gabapentin orally. Improvement was evaluated based on the neurological examination and grading. Considerable improvement was noticed after the first dose of aBM-MSCs. The animal started complete weight bearing on its pelvic limbs after two doses. CONCLUSION: Percutaneous transplantation of aBM-MSCs might have played an important role in reversing the neurological deficits secondary to IVDH in this dog. Further studies are required preferably in a larger population to confirm the efficacy of aBM-MSCs therapy in ameliorating neural deficits associated with IVDH.

Therapeutic Potential of Platelet-Rich Plasma in Canine Medicine.

Platelets are the reservoir of growth factors and play a major role in several physiological processes, such as coagulation, angiogenesis, immune response, and tissue repair. Platelet concentrates are broadly classified into two groups depending on their fibrin content, namely platelet-rich plasma (PRP) and platelet-rich fibrin (PRF). They are further divided based on their leucocyte contents. The PRP is plasma containing supra-physiological concentrations of the platelets. The growth factors present in the PRP play a crucial role in the promotion of local angiogenesis, regulation of cellular activity, stem cell homing, proliferation and differentiation of different stem cells, and deposition of matrix proteins contributing to tissue regeneration. This review aimed to establish the therapeutic potential of PRP in canine medicine with a particular focus on the applications in ophthalmology, dermatology, and musculoskeletal disorders. A systematic literature review was performed to identify the literature published during the past 20 years (2001-2021) using authentic academic databases, such as PubMed, Science Direct, Google Scholar, and Scopus. In the initial search, 556 articles were identified and based on the specific inclusion and exclusion criteria, 59 articles were selected for further analysis. The clinical efficacy of PRP depends on the number of platelets and the growth factor concentration. The PRP-based biological therapy has broad clinical applications in musculoskeletal pathologies. It is a simple, safe, and cost-effective method that can be used to treat various diseases and disorders in canine practice. For example, PRP is used for managing corneal ulcers, corneal erosion, alkali burn, keratoconjunctivitis sicca, burn wounds, chronic wounds, cutaneous ulcers, acute traumatic bone fractures, tendinopathies, cartilage pathologies, osteoarthritis, and abdominal wall defects either as monotherapy or as an adjunctive therapeutic agent. In addition, PRP is widely used as a carrier of mesenchymal stem cells for transplanting into bone defects. Therefore, allogeneic PRP therapy can be considered a simple, safe, and cost-effective method for the treatment of various diseases and disorders in canine practice. The therapeutic application of PRP in canine medicine is limited in the present study due to the lack of consensus for collection, characterization, and clinical use. Hence, further studies are required to establish the actual worth of PRP-based regenerative strategies in canine medicine.

Consecutive Hits of COVID-19 in India: The Mystery of Plummeting Cases and Current Scenario.

The novel coronavirus disease 2019 (COVID-19)-related pandemic has been in existence for almost 2 years now after its possible emergence from a wet market in the city of Wuhan of the Chinese mainland. Evidence of the emergence and transmission of this virus was attributed to bats and pangolins. The causative virus, severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2), has rapidly spread globally, affecting humans considerably with its current death toll to be over 4.7 million out of more than 233 confirmed cases as of September 2021. The virus is constantly mutating and continuously trying to establish itself in humans by increasing its transmissibility and virulence through its numerous emerging variants. Several countries have been facing multiple waves of COVID-19 outbreaks one after the other, putting the medical and healthcare establishments under tremendous stress. Although very few drugs and vaccines have been approved for emergency use, their production capabilities need to meet the needs of a huge global population. Currently, not even a quarter of the world population is vaccinated. The situation in India has worsened during the ongoing second wave with the involvement of virus variants with a rapid and huge surge in COVID-19 cases, where the scarcity of hospital infrastructure, antiviral agents, and oxygen has led to increased deaths. Recently, increased surveillance and monitoring, strengthening of medical facilities, campaigns of awareness programs, progressive vaccination drive, and high collaborative efforts have led to limiting the surge of COVID-19 cases in India to a low level. This review outlines the global status of the pandemic with special reference to the Indian scenario.

Wound Infection with Multi-Drug Resistant Clostridium Perfringens: A Case Study.

Wound infections are among public health problems worldwide. However, progress has been made in improving surgical techniques and antibiotic treatments. Misuse/overuse of antibiotics to prevent and treat bacterial infections eventually leads to increased bacterial resistance with rising incidences of multi-drug resistant (MDR) bacterial strains. The wider dissemination of antibiotics may ultimately result in ineffectiveness to antibiotic therapy, thereby complicating/graving the outcome of a patient. In the present study, a 60-year-old male patient having wound infection with MDR bacterium that ultimately required surgical amputation of the toe was investigated. For the confirmation of MDR bacterium, two culture media viz., MacConkeyAgar and Mueller Hinton Agar media were used. The sensitivity of the isolated strain for various antibiotics was tested using the disc diffusion method. The wound sample was found positive for Gram-positive bacterium that was identified as Clostridium Perfringens. The bacterium was screened for 40 antibiotics, and among all the antibiotics, it was found sensitive for only Piperacillin/Tazobactam antibiotic combination. C. perfringens bacterium caused the gas gangrene in the infected wound part of the patient. Amputation of the gangrene -affected foot part was performed by surgery, and with good medical care, the person recovered fast. To the best of our knowledge, this is the first-ever report of MDR C. perfringens single isolate harboring resistance against at least 40 antibiotics tested. More research is needed to develop really new and effective medicines that do not cross-react with antibiotics now in use and have robust activity against MDR organisms.

Recommendation and Roadmap of Mass Vaccination against Coronavirus Disease 2019 Pandemic in Bangladesh as a Lower-Middle-Income Country.

Low-income countries (LICs) and lower-middle-income countries (LMICs) are still deprived of the optimum doses of coronavirus disease 2019 (COVID-19) vaccines for their population, equal access and distribution, as well as mass immunization roadmaps to be implemented for achieving herd immunity and protection from the ongoing pandemic. In this short report, we are interacting with the world public health experts, as well as national and global leaders for warranting the mass vaccination drive to be more progressive against COVID-19 with equitable access of vaccines to LICs or LMICs to save the lives of the poorest country people and refugees. From several scientific databases, such as Google Scholar, PubMed, as well as national and international news websites, the data were collected data by utilizing appropriate keywords regarding the topic. Bangladesh might be exemplified in this brief communication as the representative of LMIC. As of October 14, 2021, 48% of the world's people have received at least one dose of the COVID-19 vaccine. In contrast, only 2.5% of people from LICs have come in under COVID-19 vaccination for at least a single shot. Both LICs and LMICs need far more vision and ambition, including political, administrative, and diplomatic progress along with enhancing the vaccination drive for their population to be immunized through simultaneous mass vaccination progress of other countries with implementing public health safety measures against the COVID-19 pandemic.

Pharmacological Effect of Caffeine on Drosophila melanogaster: A Proof-of-Concept in vivo Study for Nootropic Investigation.

A comprehensive investigation into drug candidates with nootropic activity using a proper and high throughput yet economical model organism is an important issue to consider. This proof-of-concept study was carried out to determine whether Drosophila melanogaster can be used as an in vivo screening platform to assess the nootropic activity of certain candidates for the treatment of neurodegenerative diseases. To test this, caffeine was used as a nootropic compound and a Drosophila mutant line lacking PGRP-LB with hyperactivation of NF-κB leading to early death with neurodegenerative phenotype was used as a model organism. Caffeine was orally administered via food to the PGRP-LB mutant of D. melanogaster at different concentrations (0.4 mM, 0.08 mM, 0.016 mM) prior to phenotypical observations of the survival and locomotor activity, as well as gene expression analysis, to assess the expression level of sod1, sod2, and cat genes. The results pointed out that the lifespan of D. melanogaster treated with 0.016 mM caffeine was dramatically increased; nonetheless, no changes were observed in the locomotor activity. Phenotypical analysis using a T-maze vial test demonstrated a good cognitive improvement in response to caffeine administration. Molecular analysis revealed that caffeine at a concentration of 0,016 mM induced the expression of the endogenous antioxidant genes sod1 and cat, but not sod2, signifying that the increased lifespan may be associated with a marked improvement in cytoplasmic antioxidant function. In general, the findings of the present study are in line with those previously observed in the mammalian model organism. Therefore, it can be concluded that D. melanogaster can be used as a model organism in preliminary investigation and screening of nootropic candidates prior to further testing in its mammalian counterparts.

Clinical Applications of Substance P (Neurokinin-1 Receptor) Antagonist in Canine Medicine.

Substance P binds to the Neurokinin-1 (NK-1) receptors found in the emetic center of the central nervous system (CNS) to induce emesis. Maropitant is a selective NK-1 receptor antagonist that inhibits the binding of substance P to NK-1 receptors and is commonly used to prevent and treat vomiting in dogs. This review study aimed to discuss and analyze the therapeutic potential of substance P (Neurokinin-1 receptor) antagonist with a particular focus on the drug maropitant in canine medicine. A systematic literature review was performed to identify the existing literature on the subject during the past 20 years (2001-2021) using such databases as ScienceDirect, PubMed, Scopus, and Google Scholar. The initial search identified 173 articles; however, 41 articles were selected for further analysis, based on the specific inclusion and exclusion criteria. Studies have already confirmed the role of substance P and NK-1 receptors in central pain processing, intestinal smooth muscle contraction, vasodilation, and neurogenic inflammation. Maropitant is one of the most effective veterinary antiemetic drugs that work well against peripheral and central stimuli that trigger the vomiting center. It has been already demonstrated that the therapeutic efficacy of maropitant for managing acute vomiting in dogs is associated with pancreatitis, gastritis, and parvoviral enteritis. It can also prevent and treat chemotherapy-induced emesis and delay the signs of nausea and adverse gastrointestinal effects. Regarding the broad-spectrum antiemetic activity of maropitant, it can be recommended for managing uremic vomiting in dogs. In addition, it has also exhibited an anesthetic sparing effect since the dogs treated with maropitant require a slightly lower percentage of isoflurane as an inhalational anesthetic. The NK-1 receptors are also identified in different areas of the pain pathways. Therefore, NK-1 receptor antagonists might be effective for managing visceral pain. However, further studies are required to establish the broad therapeutic potential of NK-1 receptor antagonist drugs, such as maropitant in canine medicine. It has been shown that the pain associated with the subcutaneous administration of maropitant is due to metacresol, a preservative used in some formulations. Therefore, the side effects can be eliminated by developing novel maropitant formulations specifically for dogs.

Bluetongue in India: a systematic review and meta-analysis with emphasis on diagnosis and seroprevalence.

Bluetongue (BT) is an infectious viral disease which affects a wide range of ruminants and was first reported in India in 1964. In view of the absence of comprehensive information on the BT status in India, this study presents the seroprevalence on BT in farm animals of India based-on a systematic review and meta-analysis. A systematic review was conducted to identify the published articles (2001-2018) reporting the seroprevalence of BT in sheep, goats, cattle, buffalo, camels, and Mithun (Bos frontalis) from India. From 409 research articles, 71 fulfilled the inclusion criteria and meta-analysis for proportions was carried out targeting the eligible studies. From these, 144 strata level data were extracted with a sample size of 14048 sheep, 14696 goats, 5218 cattle, 2653 buffaloes, 2062 camels, and 222 Mithun. Overall, the analyses showed that the BT seroprevalence of 43% (95% CI: 38-49%) in goats, 39% (95% CI: 33-46%) in sheep, 38% (95% CI: 25-45%) in cattle, 34% (95% CI: 20-51%) in buffaloes, 16% (95% CI: 10-22%) in camels, and 66% (95% CI: 17-95%) in Mithun. Furthermore, the meta-regression analysis suggested that serological tests, geographical region, and sample size were the prime moderators. Meta-analytic study indicates the BT seropositivity in 25.35 million sheep (95% CI: 21.5-29.9), 58 million goats (95% CI: 51.3-66.2), 66.8 million cattle (95% CI: 47.7-86), 37.0 million buffaloes (95% CI: 21.7-55.4), 0.06 million camels (95% CI: 0.04-0.09), and 0.19 million Mithun (95% CI: 0.05-0.28). The findings highlight the variation of BT seropositivity in different geographical regions of India.

Co-infection of Newcastle disease virus genotype XIII with low pathogenic avian influenza exacerbates clinical outcome of Newcastle disease in vaccinated layer poultry flocks.

Newcastle disease (ND) and avian influenza (AI) are economically important infectious diseases of poultry. Sometime, concomitant secondary viral/or bacterial infections significantly alters the pathobiology of ND and AI in poultry. As of now, the disease patterns and dynamics of co-infections caused by ND virus (NDV, genotype XIII) and Low Pathogenic AI viruses (LPAI, H9N2) are explicitly elusive. Thus, we examined the clinicopathological disease conditions due to these two economically important viruses to understand the complex disease outcomes by virus-virus interactions in vaccinated flocks. The findings of clinicopathological and molecular investigations carried on 37 commercial ND vaccinated poultry flocks revealed simultaneous circulation of NDV and AIV in same flock/bird. Further, molecular characterization of hemagglutinin (HA) and neuraminidase (NA) genes confirmed that all the identified AIVs were of low pathogenicity H9N2 subtype and fusion (F) gene analysis of detected NDVs belong to NDV class II, genotype XIII, a virulent type. The NDV and H9N2 alone or co-infected flocks (NDV + LPAI) exhibit clinical signs and lesions similar to that of virulent NDV except the degree of severity, which was higher in H9N2-NDV co-infected flocks. Additionally, avian pathogenic E. coli and mycoplasma infections were detected in majority of the ailing/dead birds from the co-infected flocks during progression of the clinical disease. Overall, the findings highlight the multi-factorial disease complexity in commercial poultry and suggest the importance of NDV genotype XIII in intensifying the clinical disease in vaccinated birds.

Cytological and immunocytological detection and differentiation of Marek's disease and lymphoid leucosis in poultry.

Marek's disease (MD) and lymphoid leucosis (LL) are the major diseases causing lymphoid tumors in chickens accounting for high economical losses. Gross examination could not yield definite diagnosis owing to their similar presentation of lesions. Thus present work was aimed for diagnosis and differentiation of MD and LL by utilizing simple cytology and novel immunocytology techniques. Cytological examination was carried out on slides with tumor touch imprints stained by simple Giemsa staining. The diagnosis was mainly achieved based on morphology of cell population. In the present study, out of a total of 595 cases examined, 502 cases had pleomorphic lymphocytic cell population suggestive of MD and 53 cases had uniform lymphocytic/lymphoblast cell population suggestive of LL, while the rest 40 cases remained inconclusive. A definitive diagnosis was achieved after performing immunocytology using specific antibodies that revealed 518 cases had reactivity for Meq oncoprotein specific for MD and 77 cases showed immunoreactivity for IgM in transformed B-cells confirming LL. The technique of immunocytology which has been useful for detecting human viral pathogens and MD in poultry has been applied for the first time as a novel, simple, rapid and inexpensive technique that could be used as an alternate test to effectively detect and differentiate MD and LL in poultry.