Receptor for Advanced Glycation End Product (RAGE) Modulates Inflammation During Feeding of the Hard Tick, Haemaphysalis longicornis in Mice.

Ticks are notorious blood-sucking ectoparasites that affect both humans and animals. They serve as a unique vector of various deadly diseases. Here, we have shown the roles of the receptor for advanced glycation end products (RAGE) during repeated infestations by the tick Haemaphysalis longicornis using RAGE-/- mice. In primary infestation, a large blood pool developed, which was flooded with numerous RBCs, especially during the rapid feeding phase of the tick both in wild-type (wt) and RAGE-/- mice. Very few inflammatory cells were detected around the zones of haemorrhage in the primary infestations. However, the number of inflammatory cells gradually increased in the subsequent tick infestations, and during the third infestations, the number of inflammatory cells reached to the highest level (350.3 ± 16.8 cells/focus). The site of attachment was totally occupied by the inflammatory cells in wt mice, whereas very few cells were detected at the ticks' biting sites in RAGE-/- mice. RAGE was highly expressed during the third infestation in wt mice. In the third infestation, infiltration of CD44+ lymphocytes, eosinophils and expression of S100A8 and S100B significantly increased at the biting sites of ticks in wt, but not in RAGE-/- mice. In addition, peripheral eosinophil counts significantly increased in wt but not in RAGE-/- mice. Taken together, our study revealed that RAGE-mediated inflammation and eosinophils played crucial roles in the tick-induced inflammatory reactions.

Zoonotic human liver flukes, a type 1 biocarcinogen, in freshwater fishes: genetic analysis and confirmation of molluscan vectors and reservoir hosts in Bangladesh.

BACKGROUND: Opisthorchiid flukes, particularly Opisthorchis viverrini, Opisthorchis felineus, Clonorchis sinensis, and Metorchis spp. are the most common fish-borne zoonotic human liver flukes (hLFs). Liver fluke infections are more prevalent in resource-deprived and underprivileged areas. We herein estimated the prevalence of the metacercariae (MC) of major hLFs in common large freshwater fishes (lFWF) marketed for human consumption from some selected areas of Bangladesh along with detection of their molluscan vectors and reservoirs. METHODS: The current status of fish-borne zoonotic hLF infections in lFWF was investigated along with their molluscan vectors and mammalian reservoir hosts in Mymensingh and Kishoreganj in Bangladesh from July 2018-June 2022 using conventional and multiple molecular techniques, such as PCR, PCR-restriction fragment length polymorphism (RFLP), sequencing, and bioinformatic analyses. The infection rate of fishes was analyzed using the Z-test and the loads of MC were compared using the chi-squared (χ2) test. RESULTS: The MC of C. sinensis, Opisthorchis spp., and Metorchis spp. were detected in 11 species of common and popular lFWF. In lFWF, the estimated prevalence was 18.7% and the mean load was 137.4 ± 149.8 MC per 100 g of fish. The prevalence was the highest (P < 0.05) in spotted snakehead fishes (Channa punctata, 63.6%). The highest rate of infection (P < 0.05) was observed with the MC of C. sinensis (11.8%). Metacercariae were almost equally (P > 0.05) distributed between the head and body of fishes. The infection rate was slightly higher in cultured (19.6%) fishes. The MC of C. sinensis, O. felineus, O. viverrini, and Metorchis orientalis in fishes were confirmed using PCR, PCR-RFLP and bioinformatics. The cercariae of opisthorchiid (Pleurolophocercus cercariae) flukes were only recovered from Bithynia spp. (3.9%, 42 out of 1089). The ova of hLFs from dogs (4.3%, 5 out of 116) and cats (6.0%, 6 out of 100), and adult flukes (M. orientalis) from ducks (41.1% 113 out of 275) were detected. CONCLUSIONS: The MC of hLFs are highly prevalent in fresh water fishes in Bangladesh. Reservoir hosts, such as street dogs, cats, and ducks carried the patent infection, and residents of Bangladesh are at risk.

Cholelithiasis in a goat associated with chronic fascioliosis in Bangladesh: A case report and review of literatures.

The objective of the present study was to describe a very rare case of gallstone (cholelithiasis) in a goat associated with chronic fasciolosis. During a routine slaughterhouse-based survey, a two-and-half-year-old female Black Bengal Goat was found to be affected with severe chronic fascioliosis characterized by the massive damage in the liver. Through systemic dissection of liver, we isolated 94 adult Fasciola spp., and by PCR, we confirmed the fluke as Fasciola gigantica. The gallbladder of the goat was oedematous. On opening the gallbladder, we recovered 255 stones of variable sizes. Stones were whitish in colour and friable, and some of the fragile stones were attached to the wall of the gallbladder. To the authors' knowledge, this is the first report of the cholelithiasis in a goat associated with F. gigantica.

Ascaridia galli, a common nematode in semiscavenging indigenous chickens in Bangladesh: epidemiology, genetic diversity, pathobiology, ex vivo culture, and anthelmintic efficacy.

Ascaridia galli is the most common nematode in chickens. Ascaridia galli is highly prevalent in chickens reared in scavenging or semiscavenging systems. Here, we studied the epidemiology, pathology, genetic diversity, ex vivo culture protocol and anthelmintic sensitivity of A. galli prevalent in indigenous chickens in Bangladesh. Through morphological study and molecular analyses, the isolated worms were confirmed as A. galli. Of the chickens examined, 45.6% (178 out of 390) were found infected. The male and young chickens were significantly (P < 0.05) more prone to A. galli infection. Prevalence of the infection was significantly (P < 0.05) lower in the summer season. In heavy infections, A. galli blocked the small intestine. Marked inflammation, increased mucus production and petechial hemorrhages were evident in the small intestine, particularly in the duodenum. Also, there were desquamation and adhesion of the mucosal villi; degeneration, necrosis of the epithelial cells and goblet cell hyperplasia. The mucosal layer was infiltrated mainly with eosinophils and heterophils. We developed a hen egg white-based long-term ex vivo culture protocol which supported the survival and reproduction of A. galli for more than a week. Levamisole (LEV) and ivermectin (IVM) efficiently killed A. galli. However, albendazole (ABZ), mebendazole (MBZ), and piperazine (PPZ) did not kill the worms even at 120 µg/mL and 1mg/mL concentrations, respectively. Taken together, our results suggest that A. galli is highly prevalent in semiscavenging chickens in Bangladesh. Ascaridia galli can be easily maintained ex vivo in egg white supplemented M199 medium. LEV and IVM, but not ABZ, MBZ and PPZ, can be used for treating and controlling A. galli infections in chickens.

Ascaridia galli infection in chicken: Pathobiology and immunological orchestra.

BACKGROUND: Ascaridia galli is the largest gut-dwelling helminth of chickens, which confers adverse effects on meat and egg production; thus, on the animal protein supply and the economy. Both adult and immature parasites affect gut health, but larval stages play a major role in pathology. AIMS: Here, we present immunology and pathology of A. galli in chickens. MATERIALS AND METHODS: Literatures were surveyed through online platforms such as PubMed, Google Scholar and Researchgate. RESULTS: The larvae cause excessive mucus production, damage to the intestinal gland, hemorrhage, anemia, diarrhea, and malnutrition. The adult worms can cause death by intestinal obstruction and intussusception. Although both cellular and humoral immunity are involved in fighting against ascariasis, the role of naturally acquired immunity is poorly defined. In cellular immunity, Th-2 cytokines (IL-4, IL-5, IL-9, and IL-13), goblet cells (mucin), gut-associated lymphoid tissues, CD8α+ intraepithelial cells, TCRγδ + T cells, and TGF-ß4 form a protective band. Type 2 immunity provides protection by forming a network of endogenous damage-associated molecular patterns, chitin, and parasitic antigens. Among antibodies, IgY is the most prominent in chickens and provides temporary humoral protection. During parasitic infection, infiltration of various immune cells is evident, especially in the intestinal epithelium, lamina propria, and crypts of the duodenum and jejunum. In chickens older than 12 weeks, gradual reduction of worm burden is more successful than the younger birds. Female chickens exert a short-lived but higher level of protection by passing IgY to chicks in the form of egg yolk antibodies. In laying conditions, immunity differs between breeds. This review provides an overview of the silent but inevitable pathological changes induced by A. galli and the interaction of host immunity with the parasite.

Food- and vector-borne parasitic zoonoses: Global burden and impacts.

Around 25% of the global population suffer from one or more parasitic infections, of which food- and vector-borne parasitic zoonotic diseases are a major concern. Additionally, zoonoses and communicable diseases, common to man and animals, are drawing increased attention worldwide. Significant changes in climatic conditions, cropping pattern, demography, food habits, increasing international travel, marketing and trade, deforestation, and urbanization play vital roles in the emergence and re-emergence of parasitic zoonoses. Although it is likely to be underestimated, the collective burden of food- and vector-borne parasitic diseases accounts for ∼60 million disability-adjusted life years (DALYs). Out of 20 neglected tropical diseases (NTDs) listed by the World Health Organization (WHO) and the Centres for Disease Control and Prevention (CDC), 13 diseases are of parasitic origin. There are about 200 zoonotic diseases of which the WHO listed eight as neglected zoonotic diseases (NZDs) in the year 2013. Out of these eight NZDs, four diseases, namely cysticercosis, hydatidosis, leishmaniasis, and trypanosomiasis, are caused by parasites. In this review, we discuss the global burden and impacts of food- and vector-borne zoonotic parasitic diseases.

Mammalian and Avian Larval Schistosomatids in Bangladesh: Molecular Characterization, Epidemiology, Molluscan Vectors, and Occurrence of Human Cercarial Dermatitis.

Schistosomiasis is a neglected tropical disease (NTD) caused by blood flukes (Schistosoma spp.). Schistosomatids affect a wide array of vertebrate hosts, including humans. In the present study, multiple species of schistosomatids were identified by isolating schistosomatid cercariae (SC) from naturally infected snails. We also described different biotic and abiotic factors influencing SC infections in snails and reported human cercarial dermatitis (HCD) for the first time in Bangladesh. A total of 22,012 snails of seven species: Lymnaea auricularia, L. luteola, Indoplanorbis exustus, Physa acuta, Viviparus bengalensis, Brotia spp., and Thiara spp., were collected and examined. Among these snails, 581 (2.6%) belonging to five species: L. luteola, L. auricularia, P. acuta, I. exustus, and V. bengalensis, were infected with SC. The rate of infection was the highest for L. luteola (11.1%), followed by L. auricularia (5.3%), and was the lowest for V. bengalensis (0.4%). Prevalence in snails was the highest in September (16.8%), followed by October (9.5%) and November (8.8%), and was the lowest in colder months, such as January (1.8%) and February (2.1%). Infections with schistosomatids were more common in larger snails and snails collected from sunny areas. We confirmed the presence of Schistosoma indicum, S. incognitum, S. nasale, S. spindale, and Trichobilharzia szidati by PCR and sequencing. Through a questionnaire survey, we detected HCD in 214 (53.5%) individuals, and the infection rate was almost equally distributed across all professions. Collectively, the present results suggest that lymnaeid snails are the main vector for Schistosoma spp. prevalent in Bangladesh, and schistosomatids with zoonotic potential are also prevalent.

Research Note: Genetic analysis, pathology, and vectors of echinostomiasis, a zoonotic helminth infection in chickens in Bangladesh.

Echinostomes (Trematoda: Echinostomatidae) are food-borne zoonotic flatworms that affect birds, animals and humans, and has been classified as neglected tropical diseases (NTDs) by the World Health Organization (WHO), which cause severe enteritis in poultry and hamper production. Here, we confirmed the species of echinostomes affecting chickens in Bangladesh along with their genetic analyses, pathology and vectors. We isolated and identified adult worms from chickens, cercariae from fresh water snails and metacerariae (MC) from some wild fishes. We recovered Echinostoma revolutum (10.3%) and Hypoderaeum conoideum (6.0%) from chickens. Zoonotic E. revolutum was confirmed by amplifying nad1 gene and subsequent sequencing. Several mutations were detected in nad1 gene and our isolates belonged to the Euro-Asian clade. We observed thickening of mucosal layer, hyperplasia of goblet cells, infiltration of eosinophils, lymphocytes and must cells in the infected intestine. About 5.3% snails were infected and the highest percentage of infection was found in Lymnaea luteola (12.1%). Echinostome infection in snails was the highest in November (9.6%) and lowest in February (3.1%) in Bangladesh. MC of echinostomes were identified from blue panchax (Aplocheilus panchax) and tank goby (Glossogobius giuris). In conclusion, echinostomiasis is a notable big problem in indigenous chickens in Bangladesh and people, especially, villagers are at risk.

Natural nodular worm infection in goats induces eosinophil extracellular DNA trap (EET) formation.

Extracellular DNA Trap (ET) formation by granulocyte is a strong innate immune machinery that plays crucial roles in trapping and killing of pathogens. Here, we show Eosinophil Extracellular DNA Trap (EET) formation in goats naturally infected with nodular worms (Oesophagostomum columbianum, Strongyloidae: Nematoda). By a slaughterhouse based survey, we found that 60% goats were infected with nodular worms. We detected numerous, hard and pale yellow to dark black nodules of variable sizes (0.25-2 cm) in the large intestine and the number of nodules were significantly (p < .05) higher in the cecum (21.7 ± 17.9) than in the colon (10.1 ± 9.9). Histologically, pink colored circumscribed caseous mass was surrounded by a dense zone of infiltration and fibrous proliferation along with massive infiltration of eosinophils in and around the necrotic mass. DAPI staining revealed huge accumulation of extracellular DNA, which formed wide ridge like structure surrounding the necrotic zone. Massive release of eosinophils cationic proteins (ECP), a helmintho-toxic substance, was found into the lesions. Collectively, our results suggest that nodular worm infection induces EETosis and ECP release, and is one of the major parasitic problem affecting Black Bengal goats that causes distortion of normal architecture of the gut wall.