Histopathological changes in the kidneys of vertebrate hosts infected naturally and experimentally with Paratanaisia bragai (Trematoda, Digenea)

Mudanças histopatológicas nos rins de hospedeiros vertebrados naturalmente e experimentalmente infectados com Paratanaisia bragai (Trematoda, Digenea)

Vanessa Barreto Xavier Aleksandra Oliveira-Menezes Marcos Antônio José dos Santos Suzana Bencke Amato Eduardo José Lopes Torres Jairo Pinheiro Solange Viana Paschoal Blanco Brandolini About the authors

Abstracts

Paratanaisia bragai is a trematode parasite that reaches sexual maturity in the kidney collecting ducts of domesticated and wild fowl and whose intermediate hosts are the snails Subulina octona and Leptinaria unilamellata. There are some discrepancies in descriptions of the pathology of this parasite in bird kidneys. Therefore, the purpose of this study was to analyze the kidneys of rock pigeons (Columba livia) naturally infected and of chickens (Gallus gallus) experimentally infected with Paratanaisia bragai, by means of macroscopic observation and by light and scanning electron microscopy. Both bird species showed significantly dilated collecting ducts. In addition, lymphocyte infiltration was observed in the kidneys of C. livia and metaplasia in the epithelial lining of the kidney collecting ducts of G. gallus.

Paratanaisia bragai ; Columba livia ; kidney; histopathology


Paratanaisia bragai é um trematódeo que atinge sua maturidade sexual nos ductos coletores de rins de aves domésticas e silvestres, tendo os moluscos Subulina octona e Leptinaria unilamellata como hospedeiros intermediários. A patologia descrita no rim das aves apresenta uma série de divergências. Dessa forma, o presente estudo teve como objetivo analisar rins de Columba livia, naturalmente infectada, e de Gallus gallus infectados experimentalmente por Paratanaisia bragai. Através das análises, verificaram-se alterações macroscópicas, por microscopia de luz e eletrônica de varredura, sendo caracterizada significativa dilatação dos túbulos coletores. Essas alterações foram observadas nas aves infectadas naturalmente e experimentalmente. Por outro lado, foi observada infiltração linfocitária nos rins de C. livia, naturalmente infectada, e ocorrência de metaplasia no revestimento epitelial dos túbulos coletores dos rins de G. gallus, experimentalmente infectados.

Paratanaisia bragai ; Columba livia ; rim; histopatologia


Introduction

Paratanaisia bragai (Santos, 1934) Freitas, 1959 is a trematode that reaches sexual maturity in the kidney collecting ducts of domesticated and wild fowls. Its main intermediate host for larval development is the snail Subulina octona (Bruguière, 1798), although there is also a report of Leptinaria unilamellata (d’Orbigny, 1837) acting as an intermediate host in Brazil (BRANDOLINI et al., 1997Brandolini SVPB, Amato SB, Pereira AA. Relacionamento de Tanaisia bragai (Digenea, Eucotylidae) e seu hospedeiro intermediár (Gastropoda, Subulinidae) sob condições experimentais. io, Subulina octonaParasitol. dia 1997; 21(3-4): 109-113.).

The embryonated eggs, which are eliminated in the host’s excretory products, passively infect the mollusk. After the miracidium hatches, two generations of sporocysts develop within the snail, cercariae and metacercariae. The definitive host acquires the infection by eating the parasitized mollusk (MALDONADO, 1945Maldonado JF. The life cycle of , Santos, 1934 (Eucotylidae), a kidney fluke of domestic pigeons. Tamerlania bragaiJ Parasitol 1945; 31(5): 306-314. http://dx.doi.org/10.2307/3273085.
http://dx.doi.org/10.2307/3273085...
; KELLER & ARAÚJO, 1992Keller GG, Araújo JLB. Ciclo evolutivo de (Santos, 1934) (Trematoda, Eucotylidae) como novo hospedeiro intermediário no Brasil: (D’Orbigny, 1835) (Gastropoda, Pulmonata, Subulinidae) em condições de laboratório. Paratanaisia bragai Leptinaria unilamellataRev Bras Parasitol Vet 1992; 1(2): 89-92.; BRANDOLINI & AMATO, 2006Brandolini SVPB, Amato SB. Desenvolvimento larval de (Santos) (Digenea, Eucotylidae) sob condições experimentais. Paratanaisia bragaiRev Bras Zool 2006; 23(4): 1097-1100. http://dx.doi.org/10.1590/S0101-81752006000400017.
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).

Infection with Paratanaisia bragai is common in poultry in the state of Rio de Janeiro (GOMES et al., 2005Gomes DC, Menezes RC, Tortelly R, Pinto RM. Pathology and first occurrence of the kidney trematode (Santos, 1934) Freitas, 1959 (Digenea: Eucotylidae) in L., 1758, from Brazil. Paratanaisia bragai Phasianus colchicusMem Inst Oswaldo Cruz 2005; 100(3): 285-288. http://dx.doi.org/10.1590/S0074-02762005000300013. PMid:16113870
http://dx.doi.org/10.1590/S0074-02762005...
). This parasite has low pathogenicity, which is indicated by the presence of mild intensity infection with no inflammatory reaction, according to Pinto et al. (2004)Pinto RM, Menezes RC, Tortelly R. Systematic and pathology study of Paratanaisia bragai (Santos, 1934) Freitas, 1959 (Digenea, Eucotylidae) infestation in ruddy ground dove Columbina talpacoti (Temminck, 1811). Arq Bras Med Vet Zootec 2004; 56(4): 472-479. http://dx.doi.org/10.1590/S0102-09352004000400008.
http://dx.doi.org/10.1590/S0102-09352004...
. However, a high parasite load can cause development of renal monostomosis with mucoid, blood, diarrhea, and even death (ARNIZAUT et al., 1992Arnizaut AB, Hayes L, Olsen GH, Torres JS, Ruiz C, Pérez-Rivera R. An epizootic of Tanaisia bragai in a captive population of Puerto Rican plain pigeon (). Columba inornata wetmoreiAnn N Y Acad Sci 1992; 653(1): 202-205. http://dx.doi.org/10.1111/j.1749-6632.1992.tb19647.x. PMid:1626872
http://dx.doi.org/10.1111/j.1749-6632.19...
).

Wild birds infected with P. bragai probably act as reservoirs because they often come into proximity by eating the same food as chickens kept outdoors (GOMES et al., 2005Gomes DC, Menezes RC, Tortelly R, Pinto RM. Pathology and first occurrence of the kidney trematode (Santos, 1934) Freitas, 1959 (Digenea: Eucotylidae) in L., 1758, from Brazil. Paratanaisia bragai Phasianus colchicusMem Inst Oswaldo Cruz 2005; 100(3): 285-288. http://dx.doi.org/10.1590/S0074-02762005000300013. PMid:16113870
http://dx.doi.org/10.1590/S0074-02762005...
). Migratory wild birds with trematode infections with can also spread helminths to new areas, representing a significant risk of native hosts being exposed to infection (ATKINSON et al., 2008Atkinson CT, Thomas NJ, Hunter DB. Parasitic diseases of wild birds. Blackell Publishing; 2008. http://dx.doi.org/10.1002/9780813804620.
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). These factors render P. bragai of veterinary importance, because infection with this parasite can cause economic losses if it results in serious lesions or death of the host.

Several authors have reported histopathological alterations caused by the presence of P. bragai in the kidney collecting tubules of domesticated and wild birds. Macroscopic changes have also been reported in Columba livia Gmelin, 1789 and Gallus gallus (Linnaeus, 1758) by Barretto & Mies Filho (1942)Barretto JF, Mies Filho AM. Primeiras observações sobre a presença de “Tamerlanea bragai” (Prof. Violantino Santos, 1934) nos rins de . Meleagris gallopavo domesticaRio de Janeiro: Departamento Nacional de Produção Animal, Serviço de Informação Agrícola, Ministério da Agricultura, Instituto de Biologia Animal; 1942. p. 3-6., in Meleagris gallopavo Linnaeus, 1758 and C. livia by Portugal et al. (1972)Portugal MASC, Oliveira GF, Fenerich FL, Cappelaro CEMPM, Chiarelli V. Ocorrência de (Santos, 1934) Freitas, 1959 (Trematoda Eucotylidae), em pomba doméstica (. Paratanaisia bragai Columbia livia domestica)Arq Inst Biol 1972; 39(3): 189-194., and in Columba inornata wetmorei Peters, 1937 by Arnizaut et al. (1992)Arnizaut AB, Hayes L, Olsen GH, Torres JS, Ruiz C, Pérez-Rivera R. An epizootic of Tanaisia bragai in a captive population of Puerto Rican plain pigeon (). Columba inornata wetmoreiAnn N Y Acad Sci 1992; 653(1): 202-205. http://dx.doi.org/10.1111/j.1749-6632.1992.tb19647.x. PMid:1626872
http://dx.doi.org/10.1111/j.1749-6632.19...
. However, several authors did not observe macroscopic alterations in all bird species infected with P. bragai, such as Menezes et al. (2001)Menezes RC, Mattos DG Jr, Tortelly R, Muniz-Pereira LC, Pinto RM, Gomes DC. Trematodes of free range reared guinea fowls (Numida meleagris Linnaeus, 1758) in the state of Rio de Janeiro, Brazil: morphology and pathology. Avian Pathol 2001; 30(3): 209-214. http://dx.doi.org/10.1080/03079450124448. PMid:19184902
http://dx.doi.org/10.1080/03079450124448...
in Numida meleagris (Linnaeus, 1758) and Pinto et al. (2004)Pinto RM, Menezes RC, Tortelly R. Systematic and pathology study of Paratanaisia bragai (Santos, 1934) Freitas, 1959 (Digenea, Eucotylidae) infestation in ruddy ground dove Columbina talpacoti (Temminck, 1811). Arq Bras Med Vet Zootec 2004; 56(4): 472-479. http://dx.doi.org/10.1590/S0102-09352004000400008.
http://dx.doi.org/10.1590/S0102-09352004...
in Columbina talpacoti (Temminck, 1811).

A few articles have reported the symptomatology resulting from infection with P. bragai. These symptoms involve weight loss, intermittent bloody mucoid diarrhea, occasional hemorrhaging, physical weakness, drowsiness and ruffled feathers (PORTUGAL et al., 1972Portugal MASC, Oliveira GF, Fenerich FL, Cappelaro CEMPM, Chiarelli V. Ocorrência de (Santos, 1934) Freitas, 1959 (Trematoda Eucotylidae), em pomba doméstica (. Paratanaisia bragai Columbia livia domestica)Arq Inst Biol 1972; 39(3): 189-194.; ARNIZAUT et al., 1992Arnizaut AB, Hayes L, Olsen GH, Torres JS, Ruiz C, Pérez-Rivera R. An epizootic of Tanaisia bragai in a captive population of Puerto Rican plain pigeon (). Columba inornata wetmoreiAnn N Y Acad Sci 1992; 653(1): 202-205. http://dx.doi.org/10.1111/j.1749-6632.1992.tb19647.x. PMid:1626872
http://dx.doi.org/10.1111/j.1749-6632.19...
; KUMAR et al., 2009Kumar BM, Taibur R, Sushanta G, Saidul I. On the incidence and pathology of dos Santos, 1934 (Freitas, 1959) infection in domestic pigeon (. Paratanaisia bragai Columba livia)J Vet Parasitol 2009; 23(2): 159-161.). Histopathological alterations have also been reported. Arnizaut et al. (1992)Arnizaut AB, Hayes L, Olsen GH, Torres JS, Ruiz C, Pérez-Rivera R. An epizootic of Tanaisia bragai in a captive population of Puerto Rican plain pigeon (). Columba inornata wetmoreiAnn N Y Acad Sci 1992; 653(1): 202-205. http://dx.doi.org/10.1111/j.1749-6632.1992.tb19647.x. PMid:1626872
http://dx.doi.org/10.1111/j.1749-6632.19...
observed interstitial infiltrate of inflammatory cells inside the renal tubules, composed of heterophils and eosinophils, while Brener et al. (2006)Brener B, Tortelly R, Menezes RC, Muniz-Pereira LC, Pinto RM. Prevalence and pathology of the nematode , the trematode , and the protozoan . Heterakis gallinarum Paratanaisia bragai Histomonas meleagridis in the turkey, Meleagris gallopavoMem Inst Oswaldo Cruz 2006; 101(6): 677-681. http://dx.doi.org/10.1590/S0074-02762006000600017. PMid:17072483
http://dx.doi.org/10.1590/S0074-02762006...
reported similar alterations, including a discrete heterophilic infiltrate.

Because of the divergent findings regarding both macroscopic and microscopic pathologies attributable to P. bragai, the aim of this study was to make a comparative analysis of the macroscopic and microscopic changes in the kidneys of C. livia naturally infected and G. gallus experimentally infected with P. bragai.

Materials and Methods

Source of the pigeons and chickens

Adult C. livia pigeons were caught in the city of Seropédica, state of Rio de Janeiro, Brazil (22° 49” 36” S and 43° 38’ 15” W) and also in the Irajá district of the city of Rio de Janeiro (22° 49’ 51” S and 43° 20” 17” W). The chickens (G. gallus, Rhode breed, 21-day-old chicks) were purchased from a breeder in the Seropédica city, located near the site where the pigeons were captured.

Experimental procedures

In the laboratory, the pigeons were subjected to fecal examination to detect infection. Four pigeons, three infected and one uninfected one (control) were euthanized in a CO2 chamber and necropsied for the removal of their kidneys, which were placed in Petri dishes containing a physiological solution of 0.85% sodium chloride (NaCl) and left there until they were sectioned by scalpel (BRANDOLINI, 2000Brandolini SVPB. Biologia de Tanaisia (Paratanaisia) bragai (Santos, 1934) (Digenea, Eucotylidae) [Tese]. Rio de Janeiro: Universidade Federal Rural do Rio de Janeiro; 2000.).

The research ethics committee of the Federal Rural University of Rio de Janeiro (UFRRJ) under protocol number 186/2011 and process number 23011596/2011-67 approved the experimental protocols.

The chicks were experimentally infected by oral administration of visceral mass from S. octona and L. unilamellata snails containing P. bragai metacercariae.

To detect the first eggs from the infected chicks, Ritchie’s fecal sedimentation method was used (as described by DE CARLI, 1994De Carli GA. Diagnóstico laboratorial das parasitoses humana: métodos e técnicas. Rio de Janeiro: MEDSI Editora Médica e Científica; 1994.), starting on the 15th day of the experiment. After confirmation of the conclusion of the prepatent period by the fecal examination, the chicks were euthanized and necropsied, following the same procedure as that used on the pigeons.

Histological procedures

Kidney fragments from both bird species were fixed in Duboscq-Brasil fluid (FERNANDES, 1949Fernandes MC. Métodos escolhidos de técnicas microscópicas. 2. ed. Rio de Janeiro: Imprensa Nacional; 1949.) and processed by the routine histological technique (HUMASON, 1979Humason GL. Animal tissue techniques. 4th ed. San Francisco: W.H. Freeman; 1979.). A Spencer 820 microtome (American Optical Co.) was used to obtain 5-µm sections, which were stained with hematoxylin-eosin and Gomori trichrome (Luna, 1968Luna LG. Manual of histologic staining methods of the armed forces institute of pathology. New York: McGraw-Hill Book Company; 1968.) and mounted on slides. Photomicrographs of the tissues were recorded using an Olympus DO12 digital image system coupled to an Olympus BX51 light microscope.

Electron microscopy of the histological slides

Tissue sections processed as described for the histological analysis were collected on coverslips, deparaffinized, gold-coated (10-15 nm thick layer) and analyzed under a FEI-Quanta 250 scanning electron microscope (SEM) operating in high vacuum mode with an acceleration voltage of 15 kV.

Results

In this study, we did not observe any symptoms in the birds parasitized by P. bragai that could be related to the infection, despite high intense infection.

In the macroscopic examination of the kidneys of C. livia naturally infected with P. bragai, some of the kidneys showed discrete dilation due to the presence of parasites in the collecting tubules. In the kidney sections, we observed a brownish coloration in the medullar region due to the presence of P. bragai inside the collecting tubules. However, the dilation observed in this study is not a recurring pattern of the macroscopic lesions observed in the kidneys of birds parasitized by P. bragai. .

The microscopic examination of the histological sections of the kidneys of naturally infected C. livia showed substantial dilation and obstruction of the renal tubules (Figure 1a), with epithelial hyperplasia due to the presence of P. bragai The kidney collecting tubules of the uninfected C. livia were also found to be lined with simple cuboidal epithelium, as can be seen in Figure 1b.

Figure 1
Kidney of Columba livia. 1a. Medullar region of a kidney naturally infected with Paratanaisia bragai, showing collecting tubules dilated by the presence of the parasite (P) and an inflammatory process (I) surrounding the tubule. Hematoxylin- eosin. Scale bar = 50 µm. 1b. Medullar region of an uninfected kidney, showing the collecting tubules (CT) with simple cuboidal epithelium. Gomori trichrome. Scale bar = 20 µm. 1c. Flattening (arrow) of the tubular epithelium (metaplasia) of a kidney from an experimentally infected G. gallus, which changed from simple cuboidal to simple squamous. Hematoxylin- eosin. Scale bar = 20 µm. 1d. Inflammatory process (I) with mononuclear cells (multifocal interstitial nephritis) in a kidney from C. livia. Gomori trichrome. Scale bar = 20 µm. 1e. Kidney collecting tubules from experimentally infected G. gallus, with formation of a digitiform structure (papilliform formation) projecting in the tubular lumen. Hematoxylin-eosin. Scale bar = 50 µm. 1f. Detail of the papilliform formation. Hematoxylin-eosin. Scale bar = 20 µm.

There was flattening of the tubular epithelium of the kidneys of the experimentally infected chicks, which changed from simple cuboidal to simple squamous (Figure 1c). An inflammatory process with mononuclear cells (multifocal interstitial nephritis) was visible in the interstitial renal tissue (Figure 1d), and also surrounding the parasitized collecting tubules, some of them showing destruction of the epithelium (Figure 1a). The collecting tubules of the parasitized chicks exhibited epithelial hyperplasia with papillary pattern projecting in the tubular lumen (Figures 1e, f), as well as proliferation of connective tissue surrounding the parasitized tubules and congestion of vessels.

The occurrence of metaplasia in the epithelium of the collecting tubule walls, from simple cuboidal to pseudostratified to prismatic, was observed in C. livia naturally infected and in G. gallus experimentally infected with the parasite (Figure 2a, b, c). The metaplasia was slightly observed in light microscopy, and the change from simple cuboidal to pseudostratified epithelium was confirmed by SEM examination of the histological sections of the infected kidneys, where nuclei of tubule wall cells of different heights were clearly visible.

Figure 2
Pseudostratified epithelium of the collecting tube wall of a kidney infected with P. bragai. 2a. Histological section of a kidney from G. gallus naturally infected with P. bragai, showing the pseudostratified epithelium with nuclei of different heights (oval detail). 2b and 2c. SEM micrographs of histological sections, showing pseudostratified epithelium with nuclei (arrows) in different positions of the collecting tubule wall after experimental infection of P. bragai (P) with eggs (E) in C. livia. Scale bars = 40 µm and 30 µm, respectively.

Discussion

The symptomatology of infection with P. bragai in various bird species has been reported in the literature, including the findings of Arnizaut et al. (1992)Arnizaut AB, Hayes L, Olsen GH, Torres JS, Ruiz C, Pérez-Rivera R. An epizootic of Tanaisia bragai in a captive population of Puerto Rican plain pigeon (). Columba inornata wetmoreiAnn N Y Acad Sci 1992; 653(1): 202-205. http://dx.doi.org/10.1111/j.1749-6632.1992.tb19647.x. PMid:1626872
http://dx.doi.org/10.1111/j.1749-6632.19...
, who observed weight loss and intermittent bloody mucoid diarrhea, resulting in the death of some animals. Likewise, Unwin et al. (2013)Unwin S, Chantrey J, Chatterton J, Aldhoun JA, Littlewood DTJ. Renal trematode infection due to in zoo housed Columbiformes and a red bird-of-paradise (. Paratanaisia bragai Paradisaea rubra)Int J Parasitol Parasites Wildl 2013; 2: 32-41. http://dx.doi.org/10.1016/j.ijppaw.2012.11.001. PMid:24533313
http://dx.doi.org/10.1016/j.ijppaw.2012....
reported the death of Zenaida graysoni Lawrence, 1871 and suggested this was caused by infection with P. bragai, because the birds exhibited severe renal lesions. Kumar et al. (2009)Kumar BM, Taibur R, Sushanta G, Saidul I. On the incidence and pathology of dos Santos, 1934 (Freitas, 1959) infection in domestic pigeon (. Paratanaisia bragai Columba livia)J Vet Parasitol 2009; 23(2): 159-161. found occasional hemorrhaging in the kidneys of C. livia parasitized by P. bragai. Other symptoms were described by Portugal et al. (1972)Portugal MASC, Oliveira GF, Fenerich FL, Cappelaro CEMPM, Chiarelli V. Ocorrência de (Santos, 1934) Freitas, 1959 (Trematoda Eucotylidae), em pomba doméstica (. Paratanaisia bragai Columbia livia domestica)Arq Inst Biol 1972; 39(3): 189-194., such as weakening, drowsiness and ruffled feathers. After these symptoms appeared, the pigeons died. The authors also described the presence of a polycystic kidney, with cysts containing a clear liquid and granular substance. Similarly, Kumar et al. (2009)Kumar BM, Taibur R, Sushanta G, Saidul I. On the incidence and pathology of dos Santos, 1934 (Freitas, 1959) infection in domestic pigeon (. Paratanaisia bragai Columba livia)J Vet Parasitol 2009; 23(2): 159-161. observed nodular growths resulting from hemorrhages in the kidneys of C. livia parasitized by P. bragai. We did not observe these symptoms, possibly because of the large dilation of the tubules, which precluded urinary obstruction (PINTO et al., 2004Pinto RM, Menezes RC, Tortelly R. Systematic and pathology study of Paratanaisia bragai (Santos, 1934) Freitas, 1959 (Digenea, Eucotylidae) infestation in ruddy ground dove Columbina talpacoti (Temminck, 1811). Arq Bras Med Vet Zootec 2004; 56(4): 472-479. http://dx.doi.org/10.1590/S0102-09352004000400008.
http://dx.doi.org/10.1590/S0102-09352004...
).

We did not observe any symptoms resulting from infection with P. bragai. Such symptoms may be related to the parasite load of the definitive host. Pinto et al. (2004)Pinto RM, Menezes RC, Tortelly R. Systematic and pathology study of Paratanaisia bragai (Santos, 1934) Freitas, 1959 (Digenea, Eucotylidae) infestation in ruddy ground dove Columbina talpacoti (Temminck, 1811). Arq Bras Med Vet Zootec 2004; 56(4): 472-479. http://dx.doi.org/10.1590/S0102-09352004000400008.
http://dx.doi.org/10.1590/S0102-09352004...
stated that high parasite loads can result in the development of certain symptomatologies and cited the symptoms reported by Arnizaut et al. (1992)Arnizaut AB, Hayes L, Olsen GH, Torres JS, Ruiz C, Pérez-Rivera R. An epizootic of Tanaisia bragai in a captive population of Puerto Rican plain pigeon (). Columba inornata wetmoreiAnn N Y Acad Sci 1992; 653(1): 202-205. http://dx.doi.org/10.1111/j.1749-6632.1992.tb19647.x. PMid:1626872
http://dx.doi.org/10.1111/j.1749-6632.19...
. Nevertheless, the severity of the microscopic lesions is not related with the parasite load, which demonstrates the low pathogenicity of P. bragai (PINTO et al., 2004Pinto RM, Menezes RC, Tortelly R. Systematic and pathology study of Paratanaisia bragai (Santos, 1934) Freitas, 1959 (Digenea, Eucotylidae) infestation in ruddy ground dove Columbina talpacoti (Temminck, 1811). Arq Bras Med Vet Zootec 2004; 56(4): 472-479. http://dx.doi.org/10.1590/S0102-09352004000400008.
http://dx.doi.org/10.1590/S0102-09352004...
). Likewise, Menezes et al. (2001)Menezes RC, Mattos DG Jr, Tortelly R, Muniz-Pereira LC, Pinto RM, Gomes DC. Trematodes of free range reared guinea fowls (Numida meleagris Linnaeus, 1758) in the state of Rio de Janeiro, Brazil: morphology and pathology. Avian Pathol 2001; 30(3): 209-214. http://dx.doi.org/10.1080/03079450124448. PMid:19184902
http://dx.doi.org/10.1080/03079450124448...
stated that despite the large size of the adult parasite, the location and average intensity of infection, these factors do not result in serious macroscopic or microscopic lesions.

However, even though very heavy infections were observed, we found symptoms that could be ascribed to the presence of P. bragai. Therefore, factors other than the parasite load should be considered, such as the conditions of the host organism, the genetic order or general order, nutritional state and existing pathologies (REY, 2008Rey L. Parasitologia. Rio de Janeiro: Guanabara Koogan; 2008.).

The findings of the macroscopic analysis in the present study corroborate those of Santos (1934)Santos V. Monostomose renal de aves doméstica. Rev Dep Nac Prod Animal 1934; 1: 203-215., who, by external examination, observed that some kidneys of C. livia and G. gallus were slightly enlarged. The same finding was reported by Barretto & Mies Filho (1942)Barretto JF, Mies Filho AM. Primeiras observações sobre a presença de “Tamerlanea bragai” (Prof. Violantino Santos, 1934) nos rins de . Meleagris gallopavo domesticaRio de Janeiro: Departamento Nacional de Produção Animal, Serviço de Informação Agrícola, Ministério da Agricultura, Instituto de Biologia Animal; 1942. p. 3-6. in M. gallopavo, by Portugal et al. (1972)Portugal MASC, Oliveira GF, Fenerich FL, Cappelaro CEMPM, Chiarelli V. Ocorrência de (Santos, 1934) Freitas, 1959 (Trematoda Eucotylidae), em pomba doméstica (. Paratanaisia bragai Columbia livia domestica)Arq Inst Biol 1972; 39(3): 189-194. in C. livia, by Arnizaut et al. (1992)Arnizaut AB, Hayes L, Olsen GH, Torres JS, Ruiz C, Pérez-Rivera R. An epizootic of Tanaisia bragai in a captive population of Puerto Rican plain pigeon (). Columba inornata wetmoreiAnn N Y Acad Sci 1992; 653(1): 202-205. http://dx.doi.org/10.1111/j.1749-6632.1992.tb19647.x. PMid:1626872
http://dx.doi.org/10.1111/j.1749-6632.19...
in C. inornata wetmorei, and by Brandolini (2000)Brandolini SVPB. Biologia de Tanaisia (Paratanaisia) bragai (Santos, 1934) (Digenea, Eucotylidae) [Tese]. Rio de Janeiro: Universidade Federal Rural do Rio de Janeiro; 2000. in C. livia. However, this finding was not reported by Menezes et al. (2001)Menezes RC, Mattos DG Jr, Tortelly R, Muniz-Pereira LC, Pinto RM, Gomes DC. Trematodes of free range reared guinea fowls (Numida meleagris Linnaeus, 1758) in the state of Rio de Janeiro, Brazil: morphology and pathology. Avian Pathol 2001; 30(3): 209-214. http://dx.doi.org/10.1080/03079450124448. PMid:19184902
http://dx.doi.org/10.1080/03079450124448...
in N. meleagris, by Pinto et al. (2004)Pinto RM, Menezes RC, Tortelly R. Systematic and pathology study of Paratanaisia bragai (Santos, 1934) Freitas, 1959 (Digenea, Eucotylidae) infestation in ruddy ground dove Columbina talpacoti (Temminck, 1811). Arq Bras Med Vet Zootec 2004; 56(4): 472-479. http://dx.doi.org/10.1590/S0102-09352004000400008.
http://dx.doi.org/10.1590/S0102-09352004...
in C. talpacoti, or by Brener et al. (2006)Brener B, Tortelly R, Menezes RC, Muniz-Pereira LC, Pinto RM. Prevalence and pathology of the nematode , the trematode , and the protozoan . Heterakis gallinarum Paratanaisia bragai Histomonas meleagridis in the turkey, Meleagris gallopavoMem Inst Oswaldo Cruz 2006; 101(6): 677-681. http://dx.doi.org/10.1590/S0074-02762006000600017. PMid:17072483
http://dx.doi.org/10.1590/S0074-02762006...
in M. gallopavo. Barretto & Mies Filho (1942)Barretto JF, Mies Filho AM. Primeiras observações sobre a presença de “Tamerlanea bragai” (Prof. Violantino Santos, 1934) nos rins de . Meleagris gallopavo domesticaRio de Janeiro: Departamento Nacional de Produção Animal, Serviço de Informação Agrícola, Ministério da Agricultura, Instituto de Biologia Animal; 1942. p. 3-6. also observed accentuated atrophy of one of the kidneys, although this was not found in the present study.

In the macroscopic examination of the kidney sections from infected birds, we observed the presence of adult parasites in the collecting tubules, as evidenced by their dark brown color, due to the large number of eggs inside their uteruses. High infection rates can cause obstruction and hypertrophy of the tubules, as observed in this study and by other authors (BARRETTO & MIES FILHO, 1942Barretto JF, Mies Filho AM. Primeiras observações sobre a presença de “Tamerlanea bragai” (Prof. Violantino Santos, 1934) nos rins de . Meleagris gallopavo domesticaRio de Janeiro: Departamento Nacional de Produção Animal, Serviço de Informação Agrícola, Ministério da Agricultura, Instituto de Biologia Animal; 1942. p. 3-6.; BRANDOLINI, 2000Brandolini SVPB. Biologia de Tanaisia (Paratanaisia) bragai (Santos, 1934) (Digenea, Eucotylidae) [Tese]. Rio de Janeiro: Universidade Federal Rural do Rio de Janeiro; 2000.; BRENER et al., 2006Brener B, Tortelly R, Menezes RC, Muniz-Pereira LC, Pinto RM. Prevalence and pathology of the nematode , the trematode , and the protozoan . Heterakis gallinarum Paratanaisia bragai Histomonas meleagridis in the turkey, Meleagris gallopavoMem Inst Oswaldo Cruz 2006; 101(6): 677-681. http://dx.doi.org/10.1590/S0074-02762006000600017. PMid:17072483
http://dx.doi.org/10.1590/S0074-02762006...
; MENEZES et al., 2001Menezes RC, Mattos DG Jr, Tortelly R, Muniz-Pereira LC, Pinto RM, Gomes DC. Trematodes of free range reared guinea fowls (Numida meleagris Linnaeus, 1758) in the state of Rio de Janeiro, Brazil: morphology and pathology. Avian Pathol 2001; 30(3): 209-214. http://dx.doi.org/10.1080/03079450124448. PMid:19184902
http://dx.doi.org/10.1080/03079450124448...
; PINTO et al., 2004Pinto RM, Menezes RC, Tortelly R. Systematic and pathology study of Paratanaisia bragai (Santos, 1934) Freitas, 1959 (Digenea, Eucotylidae) infestation in ruddy ground dove Columbina talpacoti (Temminck, 1811). Arq Bras Med Vet Zootec 2004; 56(4): 472-479. http://dx.doi.org/10.1590/S0102-09352004000400008.
http://dx.doi.org/10.1590/S0102-09352004...
; SANTOS, 1934Santos V. Monostomose renal de aves doméstica. Rev Dep Nac Prod Animal 1934; 1: 203-215.).

Based on the microscopic examination of kidney tissue sections, our observation of dilation of the collecting tubules of the medullar region due to the presence of P. bragai is in line with the findings of Santos (1934)Santos V. Monostomose renal de aves doméstica. Rev Dep Nac Prod Animal 1934; 1: 203-215., Barretto & Mies Filho (1942)Barretto JF, Mies Filho AM. Primeiras observações sobre a presença de “Tamerlanea bragai” (Prof. Violantino Santos, 1934) nos rins de . Meleagris gallopavo domesticaRio de Janeiro: Departamento Nacional de Produção Animal, Serviço de Informação Agrícola, Ministério da Agricultura, Instituto de Biologia Animal; 1942. p. 3-6., Kumar et al. (2009)Kumar BM, Taibur R, Sushanta G, Saidul I. On the incidence and pathology of dos Santos, 1934 (Freitas, 1959) infection in domestic pigeon (. Paratanaisia bragai Columba livia)J Vet Parasitol 2009; 23(2): 159-161. and Unwin et al. (2013)Unwin S, Chantrey J, Chatterton J, Aldhoun JA, Littlewood DTJ. Renal trematode infection due to in zoo housed Columbiformes and a red bird-of-paradise (. Paratanaisia bragai Paradisaea rubra)Int J Parasitol Parasites Wildl 2013; 2: 32-41. http://dx.doi.org/10.1016/j.ijppaw.2012.11.001. PMid:24533313
http://dx.doi.org/10.1016/j.ijppaw.2012....
, who analyzed Paradisaea rubra Daudin, 1800 (Passeriformes) and Z. graysoni (Columbiformes).

About the classification of the epithelium of the collecting tubule wall in the presence of P. bragai, there are some divergences in the literature, such as: Santos (1934)Santos V. Monostomose renal de aves doméstica. Rev Dep Nac Prod Animal 1934; 1: 203-215., the collecting tubules had thick walls and multistratified epithelium compressing adjacent tubules. The expression “multistratified epithelium” was also used by Ross et al. (1993)Ross MH, Reith EJ, Romrell LJ. Histologia, texto e atlas. 2. ed. São Paulo: Panamericana; 1993. to classify the epidermis, one of the layers of the skin. This term designates a lining that has various layers of flattened and juxtaposed cells.

The epithelium of the collecting tubule wall of uninfected C. livia is classified as simple cuboidal, but its appearance changes when infected, as described here, from simple cuboidal to pseudostratified and prismatic, as seen in some tubules, while in others we observed flattening of the tubule wall cells. According to Junqueira & Carneiro (1995)Junqueira LC, Carneiro J. Histologia básica. 8. ed. Rio de Janeiro: Guanabara Koogan; 1995., pseudostratified is a term employed to classify the epithelium when, although there is only one layer, it appears to have multiple layers due to the location of nuclei at different heights.

The appearance of various cell layers could explain the classification “multistratified epithelium” adopted by Santos (1934)Santos V. Monostomose renal de aves doméstica. Rev Dep Nac Prod Animal 1934; 1: 203-215.. However, the classification adopted by Santos (1934)Santos V. Monostomose renal de aves doméstica. Rev Dep Nac Prod Animal 1934; 1: 203-215. could also be mistaken, since metaplasia could have occurred in the epithelium of the collecting tubules due to infection with P. bragai, a supposition supported by the present study. The infection with P. bragai could have caused the nuclei to occupy different positions, thus making ‘pseudostratified’ rather than ‘multistratified’ the correct description. Therefore, Santos (1934)Santos V. Monostomose renal de aves doméstica. Rev Dep Nac Prod Animal 1934; 1: 203-215. results may not have been multiple layers of cells, but rather a shift in the positions of some nuclei and elongation of the cells, as we found in this study.

Flattened cells of the collecting tubule wall of experimentally infected G. gallus were also observed by Pinto et al. (2004)Pinto RM, Menezes RC, Tortelly R. Systematic and pathology study of Paratanaisia bragai (Santos, 1934) Freitas, 1959 (Digenea, Eucotylidae) infestation in ruddy ground dove Columbina talpacoti (Temminck, 1811). Arq Bras Med Vet Zootec 2004; 56(4): 472-479. http://dx.doi.org/10.1590/S0102-09352004000400008.
http://dx.doi.org/10.1590/S0102-09352004...
and Gomes et al. (2005)Gomes DC, Menezes RC, Tortelly R, Pinto RM. Pathology and first occurrence of the kidney trematode (Santos, 1934) Freitas, 1959 (Digenea: Eucotylidae) in L., 1758, from Brazil. Paratanaisia bragai Phasianus colchicusMem Inst Oswaldo Cruz 2005; 100(3): 285-288. http://dx.doi.org/10.1590/S0074-02762005000300013. PMid:16113870
http://dx.doi.org/10.1590/S0074-02762005...
, while Kumar et al. (2009)Kumar BM, Taibur R, Sushanta G, Saidul I. On the incidence and pathology of dos Santos, 1934 (Freitas, 1959) infection in domestic pigeon (. Paratanaisia bragai Columba livia)J Vet Parasitol 2009; 23(2): 159-161. found atrophy of the epithelial lining.

Arnizaut et al. (1992)Arnizaut AB, Hayes L, Olsen GH, Torres JS, Ruiz C, Pérez-Rivera R. An epizootic of Tanaisia bragai in a captive population of Puerto Rican plain pigeon (). Columba inornata wetmoreiAnn N Y Acad Sci 1992; 653(1): 202-205. http://dx.doi.org/10.1111/j.1749-6632.1992.tb19647.x. PMid:1626872
http://dx.doi.org/10.1111/j.1749-6632.19...
observed interstitial infiltrate of inflammatory cells inside the renal tubules, composed of heterophils and eosinophils, feature not observed in this study. The occurrence of inflammation near the collecting tubules as a result of infection with P. bragai observed in this study agrees with the findings of Menezes et al. (2001)Menezes RC, Mattos DG Jr, Tortelly R, Muniz-Pereira LC, Pinto RM, Gomes DC. Trematodes of free range reared guinea fowls (Numida meleagris Linnaeus, 1758) in the state of Rio de Janeiro, Brazil: morphology and pathology. Avian Pathol 2001; 30(3): 209-214. http://dx.doi.org/10.1080/03079450124448. PMid:19184902
http://dx.doi.org/10.1080/03079450124448...
, who noted discrete inflammation with the presence of heterophils around the tubules in N. meleagris parasitized by P. bragai, and with those of Silva et al. (2005)Silva MEM, Mattos-Júnior DG, Tortelly R, Menezes RC. Lesões causadas por alguns helmintos em galinha-d’angola (, L.) procedentes de estado do Rio de Janeiro. Numida meleagrisR Bras Ci Vet 2005; 12(1-3): 118-123., who observed chronic interstitial nephritis, mainly heterophilic in nature. Unwin et al. (2013)Unwin S, Chantrey J, Chatterton J, Aldhoun JA, Littlewood DTJ. Renal trematode infection due to in zoo housed Columbiformes and a red bird-of-paradise (. Paratanaisia bragai Paradisaea rubra)Int J Parasitol Parasites Wildl 2013; 2: 32-41. http://dx.doi.org/10.1016/j.ijppaw.2012.11.001. PMid:24533313
http://dx.doi.org/10.1016/j.ijppaw.2012....
also observed interstitial nephritis, while Gomes et al. (2005)Gomes DC, Menezes RC, Tortelly R, Pinto RM. Pathology and first occurrence of the kidney trematode (Santos, 1934) Freitas, 1959 (Digenea: Eucotylidae) in L., 1758, from Brazil. Paratanaisia bragai Phasianus colchicusMem Inst Oswaldo Cruz 2005; 100(3): 285-288. http://dx.doi.org/10.1590/S0074-02762005000300013. PMid:16113870
http://dx.doi.org/10.1590/S0074-02762005...
found an inflammatory reaction with granulocytes, and Brener et al. (2006)Brener B, Tortelly R, Menezes RC, Muniz-Pereira LC, Pinto RM. Prevalence and pathology of the nematode , the trematode , and the protozoan . Heterakis gallinarum Paratanaisia bragai Histomonas meleagridis in the turkey, Meleagris gallopavoMem Inst Oswaldo Cruz 2006; 101(6): 677-681. http://dx.doi.org/10.1590/S0074-02762006000600017. PMid:17072483
http://dx.doi.org/10.1590/S0074-02762006...
observed a discrete heterophilic infiltrate between mononucleated cells.

Conclusions

Infection with P. bragai can cause dilation of the renal lobes of C. livia. The presence of the parasite caused substantial dilation of the kidney collecting tubules of the birds infected naturally and experimentally.

The occurrence of lymphoplasmacytic infiltration was observed only in natural infections.

The presence of P. bragai led to metaplasia of the epithelial lining of the kidney collecting tubules in experimentally infected G. gallus.

Acknowledgements

The authors gratefully acknowledge CAPES (Brazil’s Federal Agency for the Support and Improvement of Higher Education) for its financial support, and the staff of the Otto Wucherer Helminth Biology Laboratory of the Carlos Chagas Filho Biophysics Institute, Federal University of Rio de Janeiro for their assistance with the photomicrographs.

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Publication Dates

  • Publication in this collection
    Apr-Jun 2015

History

  • Received
    01 Dec 2014
  • Accepted
    06 Feb 2015
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