Evaluation of follicular lymphoid depletion in the Bursa of Fabricius: an alternative methodology using digital image analysis and artificial neural networks

Fifty Bursa of Fabricius (BF) were examined by conventional optical microscopy and digital images were acquired and processed using Matlab® 6.5 software. The Artificial Neuronal Network (ANN) was generated using Neuroshell® Classifier software and the optical and digital data were compared. The ANN was able to make a comparable classification of digital and optical scores. The use of ANN was able to classify correctly the majority of the follicles, reaching sensibility and specificity of 89% and 96%, respectively. When the follicles were scored and grouped in a binary fashion the sensibility increased to 90% and obtained the maximum value for the specificity of 92%. These results demonstrate that the use of digital image analysis and ANN is a useful tool for the pathological classification of the BF lymphoid depletion. In addition it provides objective results that allow measuring the dimension of the error in the diagnosis and classification therefore making comparison between databases feasible.

Evaluation of bursal lymphoid depletion: comparison between the conventional histology method and digital lymphocyte depletion evaluation system

Abstract: Fifty-five bursa of Fabricius (BF) were evaluated by optical microscopy for three different avian histopathologists (H1, H3 and H4) to determine the degree of lymphoid depletion. One histologist evaluated the same slides at two different times (H1 and H2) with four-months interval between the observations. The same BFs were evaluated using the system of Digital Lymphocyte Depletion Evaluation (ADDL), being performed by three differents operators of the system, not histopathologists. The results showed was a significant difference between the histopathologists and between the scores established by the same expert (H1 and H2). However, there were not significant differences between the scores with the ADDL system, obtained using ADDL. The results make clear the fragility of the subjective lymphocyte depletion score classification by the traditional histologic method, while the ADDL system proves to be more appropriated for the assessment of the lymphoid loss in the BF.

Ação trófica de aditivos fitogênicos, glutamina e ácido glutâmico sobre a Bursa de Fabrícius e intestino delgado de frangos de corte

Abstract The aim of this study was to evaluate the effect of phytogenic additives and glutamine plus glutamic acid, associated or not, on histomorphometry of bursa of Fabricius and small intestine, oocyst count and lesion scores, and carbon turnover of duodenal mucosa of broiler chickens infected with Eimeria acervulina. A total of 450 male broiler chickens was distributed into a completely randomized design with six treatments and three replications. Treatments consisted of control diet (CD); CD + coccidiosis vaccine; CD + antibiotic performance enhancers and anticoccidial (APE/AC); CD + glutamine and glutamic acid (Gln/Glu); CD + phytogenic additives (PA); CD + Gln/Glu + PA. Birds on treatment CD + vaccine were vaccinated via drinking water at three days of age against coccidiosis. At 16 days of age all birds of all treatments were inoculated orally and individually with 500,000 oocysts of Eimeria acervulina. There was no treatment effect on lesion score in the intestinal epithelium of birds. The smaller number of excreted oocysts was observed in groups of birds fed diets containing APE/AC and PA. Were observed better results of villus height and crypt depth for duodenum and ileum of birds of treatments containing Gln/Glu at 7 days of age, and Gln/Glu and PA at 21 days of age. Higher percentage of cortical area from bursa follicles was observed in birds fed diets supplemented with Gln/Glu and PA at 7, 14 and 21 days of age. Increased turnover of intestinal mucosa was observed in treatments containing Gln/Glu, indicating acceleration in development and regeneration of damaged tissue. Glutamine plus glutamic acid and phytogenic additives can provide improvements to structure, and thus to intestinal function, as well as to better immune response against the infectious challenges. Phytogenic additives can be used for coccidiosis control of broiler chickens where the use of antibiotic performance enhancers and anticoccidials is prohibited.

Western blot detection of infectious bursal disease virus infection

In order to evaluate the use of a Western blot methodology for the diagnosis of infectious bursal disease virus (IBDV) infection, chickens were experimentally infected with IBDV strains and tested for the presence of viral antigens and antibodies by a blocking Western blot test (bWB). The viral proteins obtained from the bursa of Fabricius (BF) were transferred to a nitrocellulose membrane after sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and the chicken sera obtained by heart puncture were used for the detection of these proteins. In order to eliminate nonspecific reactions, we used a rabbit anti-chicken serum (blocking tool). By the use of the bWB test, two distinct viral proteins of 43-kDa (VP2) and 32-kDa (VP3) were detected. We suggest the use of this methodology for the detection of IBDV infection in animals suspected of having IBDV reinfection and a chronic subclinical form of the disease. With the use of the rabbit anti-chicken sera for blocking, this method is practical, sensitive and less time consuming

Diagnóstico imuno-histoquímico e caracterização anatomopatológica de clamidiose em psitacídeos

A clamidiose é causada por Chlamydophila psittaci e representa uma das principais zoonoses de origem aviária. Realizou-se um estudo retrospectivo em psitacídeos do período de 1995 a 2012 e exame imuno-histoquímico (IHQ) anti-Chlamydia. Foram avaliados 111 casos, dos quais 12 foram a óbito devido à clamidiose. As aves eram provenientes de apreensão ou cativeiro (zoológicos, criatórios, centros de triagem e domicílios). À necropsia observou-se fígado aumentado (4/12) com áreas branco-amareladas (3/12), baço aumentado (2/12) e rompido (1/12), saco pericárdico com deposição de fibrina (1/12), polisserosite fibrinosa (1/12) e em três casos não havia lesões. Na avaliação histopatológica evidenciou-se hepatite necrótica mononuclear (7/12), hepatite mononuclear (3/12), hiperplasia de ductos biliares (8/12), esplenite necrótica histiocitária (9/12), hemossiderose em fígado (9/12) e baço (9/12), aerossaculite mononuclear (4/12), pericardite fibrino-heterofílica (2/12), necrose (1/12) e rarefação (1/12) linfoide de bursa de Fabricius, pneumonia fibrinosa (1/12), nefrite mononuclear (1/12) e granulomas renais (1/12). Observaram-se inclusões basofílicas intracitoplasmáticas (corpos elementares) em fígado (2/12), baço e rins (1/12). Evidenciou-se imunomarcação anti-Chlamydia em fígado (11/12), baço (7/9), pulmões (3/9), rins (2/8), intestinos (2/3), sacos aéreos (1/4) e bursa de Fabricius (1/2). A IHQ poderá ser utilizada como forma de diagnóstico definitivo post mortem de clamidiose em psitacídeos no Brasil.

Study of chelicerae in the tick Amblyomma cajennense (Fabricius 1787) by scanning electron microscopy

Chelicerae de Amblyomma cajennense foram examinados em microscopia de varredura. Esta técnica mostrou detalhes não evidenciáveis ao poder de resolução do microscópio ótico.

Preliminary screening of the larvicidal effect of Brevibacillus laterosporus strains against the blowfly Chrysomya megacephala(Fabricius, 1794) (Diptera: Calliphoridae)

AbstractINTRODUCTION: This study evaluated whether different strains of Brevibacillus laterosporus could be used to control larvae of the blowfly Chrysomya megacephala , a pest that affects both human and animal health.METHODS:Mortality rates were recorded after 1-mL suspensions of sporulated cells of 14 different strains of B. laterosporus were added to 2.5g of premixed diet consisting of rotting ground beef fed to first instar larvae of C. megacephala . All bioassays were performed using 10 larvae per strain, with a minimum of three replicates for each bioassay. Larval mortality was recorded daily up to seven days.RESULTS:Strains Bon 707, IGM 16-92, and Shi 3 showed the highest toxicity toward the larvae producing 70.5%, 64.5%, and 51.6% of larval mortality, respectively, which was significantly higher than that in the control group (p < 0.05). In contrast, strains NRS 1642, NRS 661, NRS 590 BL 856, NRS 342, ATCC 6457, Bon 712, and NRS 1247 showed limited or no pathogenic activity against the target larvae.CONCLUSIONS:Our preliminary data indicated that B. laterosporus could be used to develop bioinsecticides against C. megacephala .

Structure of the salivary glands of the unfed female tick Amblyomma cajennense (Fabricius) (Acarina: Ixodidae)

Acini in in the salivary glands of female tick specimens of Amblyomma ajennense unfed at both postnymphal and adult phases, were studied. The salivary glands are consisted by three acini, one agranular and two granular. The agranular acini are directly attached to the anterior portion of the main salivary duct, consisting of cells without valve. A relatively large, clear, central cell occupies most of the alveolar midsection. The central cell is in contact with the acini lumen. Granular acini consist of approximately seven to fourteen cells (type II acini) or seven to sixteen (type III acini). The type II acini have three types of granular cells ("a", "b" and "c") and valve; the type III acini have another three types of granular cells ("d", "e" and "f") also presenting a valve.

Structure of the salivary glands of the unfed male tick Amblyomma cajennense (Fabricius) (Acarina: Ixodidae)

Acini in the salivary glands of unfed male Amblyomma cajennense of different ages, were studied. The salivary glands consist of one agranular and three granular acini types. The agranular acini are directly attached to the medial and anterior portion of the main salivary duct, and to some branches of the secondary ducts. A large, clear, central cell occupies the centre and this cell is in contact with the acinar lumen. There is no valve to the lumen. Granular acini consist of approximately six to fourteen cells (type II acini) or eight to thirteen (type III acini). The type II acini have three types of granular cells ("a", "b" and "c") and a valve: the type III acini have three types of granular cells ("d", "e" and "f" and a valve.

Artificial Feeding of Amblyomma cajennense (Fabricius, 1787) (Acari: Ixodidae) through Silicone Membrane

An artificial feeding system was used where citrated bovine blood was offerred to male and female Amblyomma cajennense. Vestiges of blood, sweat, hair and exfoliated skin were used as phago-stimulants placed on the surface of the silicone membrane. The ticks were collected, as engorged nymphs, from naturally infested equines, with the ecdysis occurring in the laboratory. Four hundred ticks were used, 50% being female, at three to four weeks post-ecdysis. Vestiges of blood on the silicone membrane were the most efficient phago-stimulant and the association of vestiges of blood and sweat residue smears yielded better results compared to the other phago-stimulants used

Host Specificity of Amblyomma cajennense (Fabricius, 1787) (Acari: Ixodidae) with Comments on the Drop-off Rhythm

The parasitic specificity of larval, nymph and adult Amblyomma cajennense on six different host species: Oryctolagus cuniculus, Rattus norvegicus, Gallus gallus domesticus, Anas platyrhynchus, Coturnix coturnix and Streptopelia decorata is described. In terms of the numbers of larvae and nymphs recovered, O. cuniculus was the best host species. The modal day for drop-off of larvae and nymphs was day three for the mammal hosts, but variable in the birds. We conclude that adult A. cajennense have a strong degree of specificity due to the fact that the tick failed to complete its life cycle on any of the evaluated hosts. The immature stages, on the other hand, showed a low level of specificity, most especially in the larval stage, indicating the existence of secondary hosts which probably serve as dispersers in the wild. The results also indicated a variable drop-off rhythm for larvae and nymphs in two periods, diurnal (6-18 hr) and nocturnal (18-6 hr), which differed depending upon the host.