Cross sectional epidemiological investigation on the prevalence of gastrointestinal helminths in free range chickens in Narsingdi district, Bangladesh.

Rural poultry production in Bangladesh is mainly based on the free range or backyard poultry production system. This backyard poultry plays a vital tool for poverty alleviation as well as for empowerment of poor women of this country. However, this production system has disadvantage of susceptibility to many diseases including higher burden of parasitic infection. Therefore this cross sectional epidemiological investigation was done to determine the prevalence and distribution of gastrointestinal helminths in Narsingdi district, Bangladesh. To conduct this study a total of 150 chickens from three different villages of Narsingdi district, Bangladesh (50 chickens per village) were collected by random sampling method and killed by cervical disarticulation. Thereafter, all the chickens were necropsied and gastrointestinal tracts were examined macroscopically for the presence helminth infection. In total two nematode (Ascaridia galli, Heterakis gallinarum,) and one cestode (Raillietina spp.) were identified by post mortem examination. Raillietina spp. was detected as the most prevalent helminth species (86-92 %) followed by A. galli (70-86 %), and H. gallinarum (70-76 %) in studied villages. In some chickens petechial hemorrhage were observed in the small intestinal wall which was associated with the A. galli infection and for some birds white tiny nodules were detected in case of H. gallinarum infection. No significant difference in parasite prevalence was observed between male and female bird as well as among three studied villages (P > 0.05). We observed that most of chickens were infected with more than one species of parasites. This finding suggests that the poultry production system in rural areas of Bangladesh and the environmental conditions are very favourable for the transmission and persistence of the parasite species in rural areas of Bangladesh.

The jejunal cellular responses in chickens infected with a single dose of Ascaridia galli eggs.

This histopathological study was carried out in order to investigate the cellular response in the jejunum to Ascaridia galli during the first 7 weeks of infection. Fourty-two ISA Brown chickens (7 weeks old) were infected orally with 500 embryonated A. galli eggs each while 28 chickens were left as uninfected controls. Six infected and four control chickens were necropsied at each time point 3, 7, 10, 14, 21, 28 and 42 days post-infection (dpi). Samples for histopathology were taken from three sites of the jejunoileum. Significantly higher eosinophil counts were seen in infected chickens compared to uninfected at 3, 7, 10, 14 and 28 dpi (P < 0.01). In both groups, the initial number of mast cells was high, but this high level of mast cells remained for a longer period in the infected group compared to the control group. Significantly higher counts were thus found in the infected group at 21 (P < 0.001), 28 (P < 0.01) and 42 dpi (P < 0.05). A. galli infection induced changes in the mucosal thickness as reduced villi length at 7, 10, 14, 21 and 28 dpi and in the degree of general cellular infiltration in the lamina propria of the mucosal layer. No adult worms were seen during the experiment; therefore, A. galli larvae have elicited a moderate cellular response in the lamina propria, mainly consisting of eosinophils in the early phase and later of mast cells.

The use of genetically marked infection cohorts to study changes in establishment rates during the time course of a repeated Ascaridia galli infection in chickens.

This study investigated the changes in establishment rates during the time course of a 6 week trickle infection of chickens with Ascaridia galli at two different dose levels, using a molecular marker. To differentiate early and late infection, two different egg cohorts (haplotype a and haplotype b, genetically identified using PCR-linked restriction fragment length polymorphism on the cox1 gene of the mitochondrial DNA) were used. Cohort-specific egg batches were produced by harvesting eggs from the uteri of female worms of the specific cohort. Fifty-six 8 week old Lohmann Brown Lite chickens were divided into seven groups and the infectivity of the egg batches was compared between two groups of chickens (P=0.6). The remaining chickens were allocated to four infection regimes and one control group. Group ab100 was trickle infected for 3 weeks with 100 eggs of haplotype a (twice weekly) followed by the same dose of eggs of haplotype b for another 3 weeks. Group ba100 was treated similarly but in the opposite order (haplotype b preceding a). A similar infection regime was applied for groups ab25 and ba25 but with a lower inoculation dose (25 eggs). All of the birds in these five groups (four infected and one control) were euthanased 2 weeks after the last inoculation. It was found that in the low-dose groups both the early and late infections established equally well, whereas in the high-dose groups the early infection was recovered in a significantly (P<0.001) higher proportion of chickens than the late infection, irrespective of genetic cohorts. Moreover, relatively higher proportions of the larvae from both the early and late infections were found in the posterior section of the small intestine. This result indicates the presence of dose-dependent resistance against reinfection and this resistance seems to act by reducing the establishment of late infection and by relocating the larvae from early infection.

Population dynamics of Ascaridia galli following single infection in young chickens.

The population dynamics of Ascaridia galli was studied in 70 ISA Brown layer pullets, 42 of them were each experimentally infected with 500 embryonated A. galli eggs and 28 chickens were kept as uninfected controls. Six chickens from the infected group and 4 from the control group were necropsied at 3, 7, 10, 14, 21, 28 and 42 days post-infection (d.p.i.). The mean worm recovery varied from 11-20% of the infection dose with the highest recovery at 3 d.p.i. and the lowest at 21 and 42 d.p.i. (P < 0·05). More larvae were recovered from the intestinal wall than from the content (P < 0·0001) and intestinal content larvae were longer than those from the wall (mean length 1·6 and 1 mm, respectively, P < 0·0001). Although larvae were growing over time, a population of small-sized larvae (length < 1 mm) was recovered at all d.p.i. During the first week of infection most of the larvae were located in the anterior half of the jejunoileum but they moved posteriorly with the age of infection. Thus, a subpopulation of larvae mainly in the lumen grew with time while another subpopulation remained small and associated with the mucosa. During the infection both subpopulations moved to a more posterior localization in the gastrointestinal (GI) tract.

Ascaridia galli in chickens: intestinal localization and comparison of methods to isolate the larvae within the first week of infection.

This study was conducted to observe the localization and to compare methods for isolation of minute Ascaridia galli larvae in chicken intestine. Firstly, six 7-week-old layer pullets were orally infected with 2,000 embryonated A. galli eggs and necropsied either at 3, 5 or 7 days post infection (dpi). More than 95 % of the recovered larvae were obtained from the anterior half of the jejunoileum, suggesting this part as the initial predilection site for A. galli larvae. Secondly, the intestinal wall of one layer pullet infected with 20,000 A. galli eggs 3 days earlier was digested in pepsin-HCl for 90 min. The initial 10 min of digestion released 51 % of the totally recovered larvae and the last 30 min of continuous digestion yielded only 5 %. This indicates that the majority of larvae were located superficially in the intestinal mucosa. Thirdly, 48 7-week-old layer pullets were infected with 500 A. galli eggs and necropsied at 3 dpi to compare three different larval isolation methods from the intestinal wall, viz., EDTA incubation, agar-gel incubation and pepsin-HCl digestion, resulting in mean percentages of the recovered larvae: 14.4, 18.2 and 20.0 %, respectively (P = 0.15). As conclusion, we recommended Pepsin-HCl digestion as the method of choice for larval recovery from the intestinal wall in future population dynamics study due to high efficiency and quick and simple detection. The agar-gel method was considered to be a prerequisite for molecular and immunological investigations as the larvae were more active and fully intact.

Localization of Ascaridia galli larvae in the jejunum of chickens 3 days post infection.

The normal habitat of the parasitic stages of Ascaridia galli is in the small intestine of poultry but the exact localization is poorly understood. Therefore, a histological study was conducted in order to localize the larvae during the early phase of infection. Six layer pullets seven-week old were infected orally with 20,000 embryonated A. galli eggs each, whereas four chickens were left as un-infected controls. At necropsy 3 days after infection the first half of jejunum/ileum was divided into two equally sized sections (J1 and J2). After taking samples for histology from the middle of J1 and J2 and the junction between these determined JX, the two sections were subjected to parasitological examination. A higher number of A. galli larvae were recovered from section J2 than J1 and the majority of larvae were recovered from the most profound layers. Based on histology 144 larvae were identified and their location was noted. The highest number of larvae was observed in the JX sample as compared to J1 and J2 (P<0.001). Most of them were located in the profound crypt zone of the mucosa (51%) as compared to the other zones (P<0.05). The number of larvae was higher in the lumen (63%) compared to the epithelium (32%) and lamina propria (5%) (P<0.001). A significantly higher number of eosinophils were found in lamina propria of the infected group compared to the control group (P<0.001). This experiment clearly showed that only few larvae had penetrated the epithelium and were positioned in the lamina propria at 3 days post infection. It was far more common that the larvae were localized within the epithelium or in the lumen of the crypts. It is therefore suggested that at least in this early phase "mucosal phase" is a more appropriate term to be used for the A. galli larval localization as compared to the term "histotrophic phase" currently used in many textbooks.

Survival of first stage larvae (L1) of Angiostrongylus vasorum under various conditions of temperature and humidity.

This experiment has been conducted to evaluate the viability of Angiostrongylus vasorum L1 under different conditions of temperature and humidity. In order to assess the viability, fox faecal pellets containing first stage larvae (L1) were exposed to relative humidity (RH) 95% and 75%, and to different temperatures (5°C, 18°C) and at fluctuating conditions ranging from -5°C to +5°C. Moreover, larval viability under outdoor conditions in April was also observed. Survival of the larvae was strongly influenced by temperature; however, humidity did not show any significant influence on viability. In controlled condition, 100% of the larvae were found motile and active; whereas, around 14% and 19% of the larvae kept at 5°C remained viable at RH 95% and RH 75%, respectively after 78 h. All of the larvae kept at 18°C died after 66 h. In outdoor condition, larval survivability reduced to 18% after 66 h, and in fluctuation temperature viable larvae were observed in both control and experimental conditions (15%) after 78 h.

Angiostrongylus vasorum: the 'French Heartworm'.

Angiostrongylus vasorum which is commonly known as 'French heartworm' is a snail-born parasitic disease affecting the members of the Canidae family. This parasite has a cosmopolitan distribution covering tropical, subtropical and temperate regions. However, its distribution is characterised by isolated endemic foci, with only sporadic occurrences outside this areas. During the last two decades, several sporadic occurrences in old and new endemic areas have been documented by the researchers. However, the spread of infection and dynamic consequences of this parasite in final host has not been clarified yet. Therefore, this review will focus on the morphology, biology, clinical significant as well as management of this parasitic disease.

The effect of temperature and host age on the infectivity and development of Angiostrongylus vasorum in the slug Arion lusitanicus.

Experimental infection with Angiostrongylus vasorum was conducted in Iberian slugs Arion lusitanicus. Initially, different size/age groups of juvenile slugs (small, <0.5 g and medium, 0.5-1 g) were exposed to freshly isolated first-stage parasitic larvae (L1) of A. vasorum. The slugs were subsequently incubated at 5, 10 and 15 degrees C for 6 weeks. Larval development within the slugs differed significantly with temperature. At 15 degrees C, all larvae developed into the third larval stage (L3), at 10 degrees C into the second stage (L2), whereas no development was observed at 5 degrees C. The mean larval burdens were highest in the largest group of slugs and tended to increase with higher temperature. In a second experiment isolated L1 were incubated at 5, 10 and 15 degrees C for 3 and 7 days prior to infection of slugs, which then were kept for 6 weeks at 15 degrees C. The infectivity decreased significantly with the larval storage time and the mean larval burden per slug was lower at higher incubating temperature. However, all established larvae developed into infective L3. Temperature had an effect on the development of the larvae and thus an impact on transmission of the parasite as only L3 are infective to the definitive canid hosts.