Intracytoplasmic Inclusion Bodies Research Articles

Lymphocystis Disease Virus Infection in Clownfish Amphiprion ocellaris and Amphiprion clarkii in Taiwan.

Lymphocystic disease affects over 150 species of marine and freshwater fish worldwide. In this study, the lymphocystis pathogen was found in 2 (Amphiprion ocellaris and Amphiprion clarkii) of the 9 species of clownfish. Detection of lymphocystis disease virus (LCDV) was based on histopathological study, electron microscope observation of virus particles and gene sequence analysis from the MCP region. Infected A. ocellaris hosts showed sparse, multifocal, white, stiff, papilloma-like nodules on the body, skin, gills and fins; while, on A. clarkia, nodules were found on the operculum skin. Histopathologic study showed lymphocystic cells with an irregular nucleus, enlarged cytoplasm and intracytoplasmic inclusion bodies surrounded by the cell membrane. The viral particle presents virions 180-230 nm in diameter, hexagonal in shape with an inner dense nucleoid under transmission electron micrographs (TEM). From the ML polygenetic tree, the clownfish LCVD genotype was closely related to the LCDV strain from paradise fish, Macropodus opercularis (KJ408271) (pairwise distance: 92.5%) from China, then followed by the strain from Spain (GU320726 and GU320736) (pairwise distance: 90.8-90.5%), Korea (AB299163, AB212999, AB213004, and AB299164) (pairwise distance: 91.5-80.5%) and lastly Canada (GU939626) (pairwise distance: 83%). This is the first report of lymphocystis disease in A. clarkii in Taiwan.

Identification, molecular characterization, and pathological features of orf virus in sheep and goats in Punjab province, Pakistan

Orf virus (ORFV) causes an acute, contagious, skin disease of sheep and goats which is economically important. The objectives of this study were to identify ORFV and to explore its pathological and phylogenetic profiles in 350 goats and 91 sheep of 14 districts of Punjab, Pakistan, from July 2020 to July 2021. Skin scrapings (total no. of samples = 441) of suspected animals were subjected to polymerase chain reactions, phylogenetic analysis, and pathological observations. The partial length of GIF/IL-2 gene (408bp) was successfully amplified in 58/441 samples. Phylogenetic analysis of GIF/IL2 gene showed that the study isolates belonged to ORFV-cluster I, together with the viruses reported in India and China. Pakistan ORFV isolates were shared 97.6-98.7% nucleotide and 97.6-100% amino acid identities with the reference strain (NC_005336). Moreover, Chinese ORFV-isolates were detected unique multiple amino acid substitutions (F11L, Q21H, D27N, I46V, N49S, N82D, D103N, S129G) with study isolates. Naturally infected animals were anorexic, emaciated, dull, and depressed. The macroscopic lesions included multifocal to coalescing, ulceration followed by proliferative papules, pustules, and crust formation on the epidermis of gums, lips, mouth commissure, muzzles, nose, and udder. Histopathological examination revealed hyperplasia, anastomosing rete ridges formation and degenerative changes, including spongiosis and vacuolation of epidermal cells. Keratinocytes exhibited eosinophilic intracytoplasmic inclusion bodies with pyknotic and karyorrhexis nuclei. This is the first report on molecular characterization of ORFV from Pakistan, with insight into its pathogenesis and comparative analysis of pathological alterations and genetic diversity between ORFV strains reported in different geographical areas.

Identification and pathogenetic study of tilapia lake virus (TiLV) isolated from naturally diseased tilapia

As an emerging pathogen, tilapia lake virus (TiLV) has caused severe socio-economic impacts and remains a devastating factor in wild and farmed tilapia. Early diagnosis and timely reporting of TiLV are very important and necessary. In the study, a TiLV infection event in a tilapia farm is studied and reported. Naturally diseased tilapia with clinical signs such as anorexia, exophthalmia, skin abrasion and hemorrhage are collected. Multiple tissue lesions of liver, spleen and kidney are observed by histopathological analysis, such as syncytial formation, intracytoplasmic inclusion body and necrosis. No bacterial pathogens are identified from liver, spleen and kidney of the diseased tilapia, while all of them show TiLV-positive. E-11 cells are employed for TiLV isolation, and cytopathic effects are developed following incubation with supernatants of the homogenized tissues collected from the diseased tilapia. Electron micrographs of the infected cells and purified TiLV show that nearly round-shaped, enveloped viral particles (60–80 nm) can be identified. Segment 1 of the isolated TiLV has a high sequence similarity to other isolates, but phylogenetic analysis shows that it is placed in a unique cluster. Experimental infection with the isolated TiLV can cause high mortality (> 90%) in healthy Nile tilapia, with the similar clinical signs and tissue lesions to those found in naturally infected tilapia. Moreover, vacuolation and necrosis in brain, inclusion body and necrosis in heart, megalocytes and necrosis in intestine are observed in experimentally infected tilapia. Muscle, spleen, liver, intestine, brain, heart, kidney and gill of the re-infected tilapia show TiLV-positive, and liver, spleen and brain are the main target tissues. The data so far confirm a TiLV-associated disease outbreak in China, enriching the relevant information of TiLV.

Graph theoretical network analysis, in silico exploration, and validation of bioactive compounds from Cynodon dactylon as potential neuroprotective agents against α-synuclein.

Introduction: Parkinson's disease (PD) is a chronic, devastating neurodegenerative disorder marked by the death of dopaminergic neurons in the midbrain's substantia nigra pars compacta (Snpc). In alpha-synuclein (α-Syn) self-aggregation, the existence of intracytoplasmic inclusion bodies called Lewy bodies (LBs) and Lewy neurites (LNs) causes PD, which is a cause of neuronal death. Methods: The present study is aimed at finding potential bioactive compounds from Cynodon dectylon that can degrade α-Syn aggregation in the brain, through in silico molecular docking investigations. Graph theoretical network analysis was used to identify the bioactive compounds that target α-Syn and decipher their network as a graph. From the data repository, twenty-nine bioactive chemicals from C. dactylon were chosen and their structures were retrieved from Pubchem. On the basis of their docking scores and binding energies, significant compounds were chosen for future investigation. The in silico prediction of chosen compounds, and their pharmacokinetic and physicochemical parameters were utilized to confirm their drug-likeness profile. Results: During molecular docking investigation the bioactive compounds vitexin (-7.3 kcal.mol-1) and homoorientin (-7.1 kcal.mol-1) showed significant binding energy against the α-Syn target protein. A computer investigation of molecular dynamics simulation study verifies the stability of the α-Syn-ligand complex. The intermolecular interactions assessed by the dynamic conditions indicate that the bioactive compound vitexin has the potency to prevent α-Syn aggregation. Conclusion: Interestingly, the observed results indicate that vitexin is a potential lead compound against α-Syn aggregation, and in vitro and in vivo studies are warranted to confirm the promising therapeutic capability.

Early innate immune responses of Nile tilapia (Oreochromis niloticus) during tilapia lake virus (TiLV) infection

Tilapia lake virus (TiLV) poses a serious threat to global tilapia aquaculture industry, causing massive mortality and huge economic losses. In the present study, we describe an experimental challenge of Nile tilapia using tilapia lake virus with emphasis on the expression profile of immune genes IPS-1 and TLR-7. Healthy Oreochromis niloticus (Nile tilapia) fingerlings were intraperitoneally injected with 50 μl of 1×105 TCID50/ml of TiLV prepared by growing the virus in EPC cells. Infection was followed with histopathology and gene expression analysis in the main target organs viz. brain, liver and kidney at 12, 24, 48, 72 and 96 h post challenge (hpc). Microscopic examination revealed localized blood clotting in the brain cortex, syncytial cell formation and presence of putative intracytoplasmic inclusion bodies in the liver tissue. Innate immune gene expression of IPS-1 was down regulated in the liver while slightly upregulated in kidney and brain during the early infection period. The innate sensor, TLR-7 showed downregulation in liver and upregulation in the brain, which was also evident by the very little pathology noticed in the brain. Overall, the lack of upregulation of innate immune genes in immunocompetent tissue like kidney could be due to increased multiplication of the virus in these tissues by interfering in the immune signalling pathway.

Eosinophilic homogeneous intracytoplasmic inclusion bodies in acquired epidermodysplasia verruciformis-like lesions.

Eosinophilic homogeneous intracytoplasmic inclusion bodies in acquired epidermodysplasia verruciformis-like lesions.

Detection of lymphocystis disease virus (LCDV) from yellowbar angelfish (Pomacanthus maculosus Forsskål, 1775) in Iran: Histopathological and phylogenetic analysis

Lymphocystis disease (LCD) is a benign viral disease characterized by the appearance of hypertrophic fibroblastic cells in the connective tissues of fish. In December 2021, a group of yellowbar angelfish (Pomacanthus maculosus Forsskål, 1775) which had significant mortalities (10%) and showed whitish nodules on the fins and skin, were packed in water-filled plastic bags, supplied with oxygen, and transported to the Ornamental Fish Clinic, Faculty of Veterinary Medicine, University of Tehran (Tehran, Iran). Following the clinical inspection and observation of whitish clusters of nodules mostly on the fins and skin by stereomicroscopy, wet smears of skin and fins revealed numerous hypertrophic lymphocystis cells. Histopathological study of the whitish nodules in skin and fins revealed many hypertrophic cells with thick hyaline capsules, irregular nucleus, and multiple intracytoplasmic inclusion bodies. In addition to the extension of hypertrophic lymphocystis cells' growth until the superficial muscle layer, infiltration of inflammatory cells and hemorrhage between hypertrophic cells are also observed. The hyaline capsule of hypertrophic fibroblasts stained with the PAS. Molecular characterization (PCR) confirmed the presence of Lymphocystis disease virus (LCDV) in the external nodules and the internal organs homogenate of the infected fish. Sequence analysis of the MCP gene of the detected virus (IR_LCDV_RGZIS) assigned it to genogroup 5. This is the first detection and characterization of LCDV in yellowbar angelfish (Pomacanthus maculosus) in Iran.

Pathogenesis of Bohle iridovirus infection in Krefft's freshwater turtle hatchlings (Emydura macquarii krefftii).

Ranaviruses have been detected in over 12 families of reptiles including many genera of turtles, tortoises, and terrapins, but the pathogenesis of these infections is still poorly understood. Krefft's river turtle hatchlings (N = 36; Emydura macquarii krefftii) were inoculated intramuscularly with Bohle iridovirus (BIV, Ranavirus, isolate) or saline, and euthanized at 9 timepoints (3 infected and 1 control per timepoint) over a 24-day period. Samples of lung, liver, kidney, and spleen were collected for quantitative polymerase chain reaction (PCR); internal organs, skin, and oral cavity samples were fixed for histopathological examination. The earliest lesions, at 8 days postinoculation (dpi), were lymphocytic inflammation of the skin and fibrinoid necrosis of regional vessels at the site of inoculation, and mild ulcerative necrosis with lymphocytic and heterophilic inflammation in the oral, nasal, and tongue mucosae. Fibrinonecrotic foci with heterophilic inflammation were detected in spleen and gonads at 16 dpi. Multifocal hepatic necrosis, heterophilic inflammation, and occasional basophilic intracytoplasmic inclusion bodies were observed at 20 dpi, along with ulcerative lymphocytic and heterophilic tracheitis and bronchitis. Tracheitis, bronchitis, and rare bone marrow necrosis were present at 24 dpi. Of the viscera tested for ranaviral DNA by PCR, the liver and spleen had the highest viral loads throughout infection, and thus appeared to be major targets of viral replication. Testing of whole blood by qPCR was the most-effective ante-mortem method for detecting ranaviral infection compared with oral swabs. This study represents the first time-dependent pathogenesis study of a ranaviral infection in turtles.

Isolation and molecular characterization of lumpy skin disease virus from hard ticks, Rhipicephalus (Boophilus) annulatus in Egypt

BackgroundLumpy skin disease (LSD), a disease of cattle and buffaloes, has recently become widely prevalent in Egypt. The aim of this study was to ascertain the potential role of Rhipicephalus (Boophilus) annulatus ticks in the transmission of this disease. Samples collected from suspected lumpy skin disease virus (LSDV) infected cows that had previously been vaccinated with the Romanian sheep pox virus (SPPV) in various Egyptian governorates were obtained between May to November over two consecutive years, namely 2018 and 2019. Ticks were morphologically identified and the partial cytochrome oxidase subunit I gene (COI) were sequenced, revealing that they were closely related to R. (Boophilus) annulatus. The G-protein-coupled chemokine receptor (GPCR) gene of the LSDV was used to test hard ticks.ResultsTwo positive samples from Kafr El-Sheikh province and one positive sample from Al-Behera province were reported. BLAST analysis revealed that the positive samples were closely related to the Kazakhstani Kubash/KAZ/16 strain (accession number MN642592). Phylogenetic analysis revealed that the GPCR gene of the LSDV recently circulating in Egypt belongs to a global cluster of field LSDV with a nucleotide identity of 98–100%. LSDV isolation was successfully performed four days after inoculation using 9 to 11-day-old embryonated chicken eggs showing characteristic focal white pock lesions dispersed on the choriallantoic membrane after three blind passages. Intracytoplasmic inclusion bodies, cell rupture, vacuoles in cells, and virus particles ovoid in shape were demonstrated by electron microscopy.ConclusionIn this study the role of hard ticks in the transmission of the LSDV to susceptible animals in Egypt was revealed and confirmed by various methods.

Clinicopathological and molecular studies on cattle naturally infected with lumpy skin diseases in selected districts of Wolaita Zone, Southern Ethiopia

BackgroundLumpy skin disease is a contagious viral disease of cattle caused by LSDV that results in huge economic losses in the cattle industry. This study characterizes LSDV in cattle through clinicopathological and molecular techniques in selected districts of Wolaita Zone, Southern Ethiopia.MethodsA crossectional study was conducted from November 2020 to June 2021 using Real-time polymerase chain reaction and Histopathological techniques to confirm LSDV.ResultThis study revealed that the percentage of positivity of cattle for LSDV was 36.2%. Clinically, cattle infected with LSDV revealed fever (39–41 °C), nodular lesions on the skin and mucous membranes, and lymphadenopathy. Histopathologically, affected tissue revealed ballooning degenerations of the epidermis, infiltration of mononuclear inflammatory cells, vasculitis, and intracytoplasmic eosinophilic inclusion bodies. RT-PCR confirmed that DNA extracts from skin biopsies of virus isolates were positive for LSDV.ConclusionThe present study confirms that LSDV is widely circulating in cattle of selected districts of the Wolaita zone. Thus, effective control measures through regular vaccination and further confirmation of circulating strains of LSDV through detailed molecular analysis should be recommended.

Oxidative stress, biochemical, and histopathological changes associated with acute lumpy skin disease in cattle.

Background and Aim:Lumpy skin disease (LSD) is a highly infectious endemic viral disease of cattle in Africa and the Middle East. The objectives of this study were to assess histopathological changes in cattle infected with LSD and measure serum malondialdehyde (MDA – oxidant) and total anti-oxidant capacity (TAC – anti-oxidant), trace elements (zinc, copper, and iron), cytokines (interleukin [IL]-1β, IL-6, and tumor necrosis factor-alpha [TNF-α]), haptoglobin (Hp), serum amyloid A (SAA), aspartate aminotransferase (AST), alanine aminotransferase (ALT), creatine phosphokinase (CPK), blood urea nitrogen (BUN), and creatinine concentrations.Materials and Methods:Blood samples were collected from a total of sixty native and mixed breed cattle; (healthy; n = 25) and (LSD diseased; n = 35). Serum concentrations of MDA and TAC were measured by colorimetric methods. Serum IL-1β, IL-6, IL-10, TNF-α, Hp, and SAA concentrations were determined using human-specific enzyme-linked immunoassay kits.Results:Serum MDA, cytokine (IL-1β, IL-6, and TNF-α), Hp, SAA, AST, ALT, CPK, BUN, and creatinine concentrations were significantly higher, while TAC, IL-10, zinc, copper, and iron concentrations were significantly lower in LSD compared to healthy cattle (p < 0.05). Cows and exotic mixed breed cattle were at higher risk of LSD oxidative stress than bulls and local breeds (p < 0.05). Age was not associated with the risk of LSD (p > 0.05). Histologically, there was extensive tissue necrosis, severe vasculitis, mononuclear cell infiltration, and intracytoplasmic inclusion bodies.Conclusion:LSD is associated with pro-oxidative and pro-inflammatory states from imbalances that favor pro-oxidant and pro-inflammatory factors in the detriment of anti-oxidant and anti-inflammatory factors, leading to organ dysfunction and ultimately death. Oxidative stress is more frequent in cows and mixed breed cattle than in bulls and local breeds.

Laporan Kasus Terapi Kombinasi Kuretase dan &lt;i&gt;Cimetidine&lt;/i&gt; Oral pada Pasien Moluskum Kontangiosium

Background: Molluscum contagiosum is an infectious disease of the skin caused by poxvirus. The virus may be transmitted through direct skin and mucous membrane contact, including sexual contact, and through contaminated fomites. The prevalence of sexually transmitted molluscum contagiosum has increased in recent years. This disease causes anxiety and discomfort for the patients. Diagnosis of molluscum contagiosum is made based on history taking, physical examination, dermoscopic examination, and histopathological examination. Proper diagnosis and management can help to reduce risk of recurrence. Case: A 26-year-old woman with main complaint of developing shiny white rashes on her thighs and buttocks since 3 months ago. The nodules felt itchy yet painless. At first, only 4 papules appeared on the buttocks. In the last month, the nodules multiplied and spread to the thighs. The patient had tried to squeeze the nodule and rice-like white mass came out. The patient had history of unprotected sexual intercourse. Dermatological examination on the right and left femoral regions and the gluteus, found that there were discrete, multiple shiny white pearl-like papules of 0.1-0.5 cm in size, smooth surface, well-defined, dome-shaped, umbilicated with caseous plug. Dermoscopic examination showed a white to yellow polylobular appearance with crown vessel. Histopathological examination showed an appearance of intracytoplasmic eosinophil inclusion bodies (molluscum bodies). Discussion: Clinical findings such as discrete multiple, pearl-colored, umbilicated dome-shaped papules, are characteristic features of molluscum contagiosum. Dermoscopic examination results and histopathological images also support the diagnosis of molluscum contagiosum. Treatment was carried out using curettage on the lesions and prescription of oral cimetidine 2x800 mg for 2 months. No recurrence was found after 5 months of follow-up

First report on the isolation of Chlamydia abortus from female dromedary camels with ovarian hydrobursitis

This study sought to isolate Chlamydia abortus (C. abortus) from camels with ovarian hydrobursitis (OVHB). To accomplish this goal, bursal tissue (n = 5) and bursal fluid (n = 6) samples were collected from 11 female dromedary camels with unilateral OVHB. A quantitative polymerase chain reaction (qPCR) was used for the preliminary detection of C. abortus in the infected samples. For the purpose of isolation, the prepared samples were inoculated into embryonated chicken eggs. Giemsa, Gimenez, and direct immunofluorescence (DIF) staining were used to detect any chlamydial inclusions in the infected yolk sacs. A second qPCR was then performed on the infected yolk sacs. The C. abortus gene was detected in 83.8% of the infected bursal tissue and bursal fluid samples. All the yolk sac smears treated with Giemsa, Gimenez, and DIF staining revealed intracytoplasmic inclusion bodies. Moreover, hemorrhagic patches, massive congestion, macerated yolk sacs, and dwarfism were observed in the infected chicken embryos. The C. abortus gene was also found in 63.6% of the infected yolk sacs. In conclusion, this is the first report of C. abortus isolation from female dromedary camels with OVHB, which represents a key step toward developing a practical vaccine and avoiding fertility problems in female camels.

Clostridium piliforme and canine distemper virus coinfection in 2 domestic dog littermates and a gray fox kit.

Concurrent Clostridium piliforme and canine distemper virus (CDV) infection was diagnosed in 2 canine littermates and 1 gray fox kit from Texas, USA. In all 3 animals, intracytoplasmic, filamentous bacteria, consistent with C. piliforme, were present along the margins of foci of hepatic necrosis. Additional histologic findings included intracytoplasmic and intranuclear inclusion bodies in bile duct and bronchial epithelial cells of the fox kit, and mild intestinal necrosis in 1 puppy. PCR assays confirmed the presence of C. piliforme in all 3 animals, CDV in both puppies, and canine parvovirus in 1 puppy. Fluorescent antibody testing confirmed the presence of CDV in the fox kit. Concurrent canine distemper and Tyzzer disease in canine littermates and the gray fox has not been reported previously, to our knowledge.

Molecular and histopathological characterization of lumpy skin disease in cattle in northern Vietnam during the 2020-2021 outbreaks.

Lumpy skin disease (LSD) is a serious emerging infectious disease in cattle caused by a virus of the family Poxviridae. According to the Department of Animal Health, LSD first occurred in Vietnam at the end of October 2020 in Cao Bang and Lang Son provinces. So far, the disease has infected over 63,000 animals, resulting in 9170 deaths occurring in 32 different provinces in northern and central Vietnam. In this study, skin samples from lumpy skin disease virus (LSDV)-infected cattle from the northern provinces of Vietnam displaying clinical symptoms including fever (> 40°C), runny nose, drooling, and skin lesions were used for genetic characterization and histopathology. Genetic analysis of the partial P32 (LSDV074), partial F (LSDV117), complete RPO30 (LSDV035), and complete G-protein-coupled-chemokine-like receptor (GPCR) (LSDV011) genes showed that all Vietnamese LSDV strains belonged to the genus Capripoxvirus and were closely related to LSDV strains isolated in China. Microscopic examination of the skin lesions showed thickening of the epidermal layer of the skin and hair follicles, hyperplasia of sebaceous glands, intracytoplasmic inclusion bodies, and hemorrhages in the mesoderm.

An Investigation of the Cause of Wild Turkey Mortality in Mississippi.

Lymphoproliferative disease virus (LPDV) is an exogenous alpharetrovirus that sporadically causes fatal lymphoid neoplasia in affected turkeys. Previous studies of wild turkeys (Meleagridis gallopavo) in the United States have demonstrated geographically widespread LPDV infection and frequent coinfection with avian poxvirus (APV) and reticuloendotheliosis virus (REV). This study was conducted to better understand health risks to Mississippi wild turkeys, including the relative importance of LPDV, APV, and REV in contributing to mortality. Thirteen wild turkeys, which died naturally or were euthanized due to illness, were submitted to Mississippi State University's Poultry Research and Diagnostic Laboratory for postmortem examinations. Birds originated from nine counties across the state over the past 5 yr. Carcasses were submitted as fresh (nonfrozen) or frozen. At autopsy, 9 of 13 turkeys had severe, proliferative cutaneous lesions on the head and neck, with diphtheritic or proliferative oral and esophageal lesions. Samples were collected for molecular diagnostic testing (LPDV and REV PCR), histopathology, and bacterial culture and isolation. External and internal parasites were preserved in formalin for identification. APV (cutaneous and/or diphtheritic forms) was diagnosed in 9 of 13 birds by identification of pathognomonic histologic lesions (including intracytoplasmic inclusion bodies). Interestingly, all birds with APV were also REV PCR positive. Furthermore, eight turkeys were positive for LPDV, and LPDV was commonly associated with coinfections with APV and REV.

Molluscum Contagiosum Masquerading as Ocular Surface Squamous Neoplasia

A 72-year-old woman was referred with suspicion of ocular surface squamous neoplasia because 2 leukoplakic nodules were noted on the bulbar conjunctiva (Fig A). Medical history was significant for atopic keratoconjunctivitis and atopic dermatitis. The nodules were excised en bloc followed by alcohol epitheliectomy. Unexpectedly, histopathology of the conjunctival tissue (hematoxylin & eosin stain) showed a cup-shaped lesion with hyperplastic epithelium surrounding intracytoplasmic eosinophilic inclusion bodies characteristic of molluscum contagiosum (Fig B).

Molecular and pathological screening of canine distemper virus in Asiatic lions, tigers, leopards, snow leopards, clouded leopards, leopard cats, jungle cats, civet cats, fishing cat, and jaguar of different states, India.

The present investigation was conducted to rule out canine distemper (CD) diseases in Indian wild felids (Asiatic lions, tigers, leopards, snow leopards, clouded leopards, leopard cats, jungle cats, civet cats, fishing cat, and jaguar). The collected samples were screened for CD virus (CDV) by histopathology (HP), immunohistochemistry (IHC) and reverse transcriptase-polymerase chain reaction (RT-PCR) targeting H gene and N gene. The HP and IHC of suspected samples portrayed that 22 [11 leopards, 6 lions, 3 tigers, 1 snow leopard and 1 civet cat] out of 129 (17.05%) wild felids were positive for CD. The major pathological consequences were observed in spleen, lung, kidney and brain. The syncytia and intranuclear as well as intracytoplasmic eosinophilic inclusion bodies were seen in CDV infected cells. Although the histopathological lesions in spleen were more specific and consistent, however, the severe demyelinated leukoencephalitis (usually expected in CD infected dog) was not observed in the brain of any Indian wild felids. Conversely, the CDV antigen has been portrayed via IHC in pancreatic islets of Langerhans of tiger species for the first time in this study. Moreover, the concurrent CD and babesiosis has also been observed in a lioness without a usual coffee-coloured urine. The N gene and H gene of CDV isolates were amplified, sequenced and subsequently constructed the phylogenetic tree. The phylogenetic analysis of H gene revealed that the CDV isolates from Indian lion formed separate clade with CDV isolates from Indian dog and Indian palm civet cat. Furthermore, two CDV isolates from Indian tigers formed clade with Onderstepoort vaccine strain and CDV isolates from dogs of Uttar Pradesh, USA and UK. Evidently, CDV is circulating in Indian wild felids and causing diseases in them.

Histopathological findings of the nasopharynx-associated lymphoid tissue of pigs co-infected with porcine circovirus 2 and porcine reproductive and respiratory syndrome virus.

Porcine circovirus 2 (PCV2) causes porcine circovirus-associated disease, and co-infection with porcine reproductive and respiratory syndrome virus (PRRSV) severely affects the pig breeding industry. Both viruses target the macrophages in lymphoid tissues. Various porcine pathogens enter via the nasal cavity, and the nasopharynx-associated lymphoid tissue (NALT) acts as the mucosal immune system. However, the pathological analysis has not progressed. This study aimed to histologically examine the NALT of pigs with suspected PCV2 and PRRSV infections. Six pigs were subjected to necropsy, and their NALT, tonsils, and mesenteric lymph nodes were collected. Macrophages, lymphocytic depletion, multinucleated giant cells, intracytoplasmic inclusion bodies, and neutrophil infiltration increased in the NALT. In situ hybridization revealed positive signals for PCV2 in the NALT of all pigs and PRRSV in the NALT of three pigs. PCV2-positive macrophages were mainly identified in the follicles, whereas PRRSV-positive tissues were found primarily around the crypt and directly below the epithelium. Quantitative PCR revealed 108-1010 copies of PCV2 DNA/µL and 102-104 copies of PRRSV DNA/µL in the NALT. Therefore, both PCV2 and PRRSV were detected in the NALT of pigs. In conclusion, the infection and replication of both viruses in the NALT and tonsils may suppress host immunity and promote co-infection with other pathogens.

Pathology and Molecular Characterization of Diphtheritic form of Fowl Pox in Chicken: A Case Report

Fowl pox is viral disease affecting the poultry species mainly chicken, turkey and pigeons worldwide. It exists in two forms viz. cutaneous and diphtheritic form. Grossly, laryngeal opening was completely occluded by caseous material and trachea was severely congested. Microscopically, trachea revealed mucosal thickening due to the proliferation of epithelial cells with characteristic intracytoplasmic eosinophilic inclusion bodies. The collected samples were screened for the P4b gene of fowl pox through PCR. Phylogenetic analysis revealed that the current FPV is closely related to the strains isolated from India and Brazil.