Germ cell survival and development critically depend on the cells' contact with Sertoli cells in the vertebrate testis. Fish and amphibians are different from mammals in that they show a cystic type of spermatogenesis in which a single germ cell clone is enclosed by and accompanied through the different stages of spermatogenesis by an accompanying group of Sertoli cells. We show that in maturing and adult testes from African catfish and Nile tilapia, Sertoli cell proliferation occurs primarily during spermatogonial proliferation, allowing the cyst-forming Sertoli cells to provide the increasing space required by the growing germ cell clone. In this regard, coincident with a dramatic increase in cyst volume and number of germ cells per cyst, in Nile tilapia, the number of Sertoli cells per cyst was strikingly increased from primary spermatogonia to spermatocyte cysts. In both African catfish and Nile tilapia, Sertoli cell proliferation is strongly reduced when germ cells have proceeded into meiosis, and stops in postmeiotic cysts. We conclude that Sertoli cell proliferation is the primary factor responsible for the increase in testis size and sperm production observed in teleost fish. In mammals, Sertoli cell proliferation in the adult testis is not observed under natural conditions. However, on the level of the individual spermatogenic cyst--similar to mammals--Sertoli cell proliferation ceases when germ cells have entered meiosis and when tight junctions are established between Sertoli cells. This suggests that fish are valid vertebrate models for studying Sertoli cell physiology.

The main objectives of the present study were to investigate the effects of 6-n-propyl-2-thiouracil (PTU) on Sertoli cell proliferation, germ cell number, and testis size in Nile tilapias (Oreochromis niloticus). In this regard, young fish (approximately 1 g BW and approximately 3.5 cm total in length) were treated for a period of 40 d with different concentrations (100 and 150 ppm) of PTU. The animals were killed and analyzed on d 1, 30, 40, 98, and 208 after the beginning of the treatment. On d 30 and 40 the spermatogenic process was delayed in fish treated with PTU compared with the control group. Also at these periods, treated tilapia had decreased (P < 0.05) body weight and total length. On d 98 body weight and total length had recovered in PTU-treated fish and were similar (P > 0.05) to those of the controls. However, testis weight and gonadosomatic index (testis mass/body weight) were approximately 100% higher (P < 0.05) in treated tilapia. Similarly, the area occupied by seminiferous tubules, the number of Sertoli cells and germ cells per cyst, and the number of Leydig cells per testis were significantly (P < 0.05) greater in treated fish. Nevertheless, nuclear volume and individual Leydig cell volume were significantly lower (P < 0.05) in tilapia receiving PTU treatment. Compared with controls, at 208 d all parameters analyzed presented the same trend as that observed at 98 d. In general, at 98 d the different PTU concentrations used during the treatment period induced similar effects. However, at 208 d the mean values observed for several parameters were significantly higher (P < 0.05) in fish exposed to 150 ppm. Probably due to the higher density of Sertoli cells per cyst in treated tilapia, these cells presented a smaller (P < 0.05) nucleolus and a trend to decrease its support capacity (efficiency). However, the meiotic index (germ cell loss during the two meiotic divisions) was similar (P > 0.05) in the three groups of fish investigated. Remarkably, the results found in tilapia were similar to those found for rats treated with PTU. This suggests strongly that the mechanisms of control of Sertoli cell and Leydig cell proliferation seem to be preserved during vertebrate evolution.

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Clinical, gross, and histopathology lesions and molecular characterization of Trichomonas spp. infection were described in two striped owls (Asio (Rhinoptynx) clamator), one American kestrel (Falco sparverius), two green-winged saltators (Saltator similis), and in a toco toucan (Ramphastos toco) from Brazil. These birds presented clinical signs including emaciation, ruffled feathers, abundant salivation and open mouth breathing presumably due to abundant caseous material. Gross lesions were characterized by multifocal yellow friable plaques on the surface of the tongue, pharynx and/or caseous masses partially occluding the laryngeal entrance. In the owls, the caseous material extended into the mandibular muscles and invaded the sinuses of the skull. Histopathologically, marked necrotic and inflammatory lesions were associated with numerous round to oval, pale eosinophilic structures (6-10μm) with basophilic nuclei, consistent with trichomonads. Organisms similar to those described above also were found in the liver of the two green-winged saltators. To the authors' knowledge, this is the first report of trichomonosis in a striped owl and a toco toucan. Sequence analysis of the Trichomonas spp. internal transcribed spacer 1 (ITS-1) region and partial 5.8S of the ribosomal RNA (rRNA) disclosed significant genetic diversity. Two sequences had 100% identity to Trichomonas gallinae, whereas two sequences had a 99% and 92% identity to a Trichomonas vaginalis-like sequence, respectively. One sequence (green-winged saltator 502-08) had a 100% identity to a newly recognized genus Simplicomonas.

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Mycoplasma gallisepticum (Mg) infection of wild native Brazilian psittacines (Psittaciformes) which died of any cause during sorting, rehabilitation, or conservation, was investigated by PCR. Two previously described PCR methodologies using Mg specific primers were employed for the analyses of 140 swab samples (cloaca, trachea, or palatine cleft). Average positive Mg detection in cloacal swabs was 51.9%, with 80.0% (n=5) of Blue-and-yellow Macaws (Ara ararauna), 60.0% (n=3) Dusky Parrots (Pionus fuscus), 52.5% (n=59) Amazon Parrots (Amazona aestiva), 50.0% (n=2) Orange-winged Parrots (Amazona amazonica), 50.0% (n=2) Jandaya Parakeetsor Jandaya Conures (Aratinga jandaya), 0% (n=2) Golden Conures or Golden Parakeets (Guarouba guarouba), and 0% (n=2) Hyacinth Macaws (Anodorhynchus hyacinthinus). Palatine cleft swab sampling was more sensitive to detect Mg, with 85.4% (n=17) detection rate, as compared to 67.4% (n=46) obtained with tracheal samples, and 53.5% (n=77) with cloacal swabs. The surprisingly high Mg incidence in psittacines kept in conservation or triage environments is possibly due to the proximity or cohabitation with several bird species during confinement and housing psittacines of different origins together. The implementation of biosecurity measures and species-specific facilities is recommended.

The incidence of the psittacine beak and feather disease virus (BFDV) was investigated in Brazilian native parrots with normal feathering arriving at rescue and triage centers for wild animals (CETAS, IBAMA) in the state of Minas Gerais, Brazil. BFDV DNA was investigated by previously described PCR technique for the partial amplification of BFDV ORF-1 in DNA extracts from blood, cloacal swab or liver of psittacines. Some birds provided more than one sample. Nine species of psittacines were sampled between January 2009 and October 2010. Blood (n=46) or cloacal swab (n=128) samples were obtained from psittacines immediately upon arrival at the triage centers. Liver samples were collected from necropsied birds dead on arrival (n=167). All swab samples were negative, except for one Ara ararauna individual (n=3) which blood presented the BFDV DNA. On the other hand, 11 liver samples were positive for BFDV DNA, with a prevalence of 7.8% in Amazona aestiva (n=140). No BFDV DNA was detected in the liver of Amazona amazonica (n=11), A. vinacea (n=5), A. rhodochorytha (n=4), Anodorhynchus hyacinthinus (n=3), Ara ararauna, (n=3), Aratinga leucophtalma (n=2), Guarouba guarouba (n=1) and Pionus maximiliani (n=1). In most cases, alopecia was not associated with BFDV detection in liver, and liver histopathology was inconclusive. Although all cloacal swab samples were negative, a few psittacines (n=19) that died at CETAS-Belo Horizonte were retested, and 21% were detected as positive in liver. A group of psittacines (n=16) was clinically evaluated, and despite showing feather dystrophy, all birds were negative in the cloacal swabs, except for one, which blood sample was positive (A. ararauna). The obtained sequences of the BFDV strains BH 215 and BH 732 were deposited in the GenBank (JQ649409 and JQ649410). A 98% similarity with strain sequences described in Australia, Japan, and New Zealand was observed. It is possible that these strains arrived in Brazil through the legal and illegal trade of parrots. However, it was not possible to associate BFDV infection with the geographical origin of birds and no local marker was detected. The rates of detection, although similar to other studies, indicate the tendency of a high incidence of the disease, possibly associated with stress, and high bird density and wide transmission in captivity conditions.

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