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Blood and fecal samples of chukar partridge (Alectoris chukar), albino pheasant (Phasianus colchicus), silver pheasant (Lophura nycthemera), rose-ringed parakeet (Psittacula krameri) and turkeys (Meleagris gallopavo) were analyzed to check parasitic prevalence. To record parasites these five avian species were placed kept in separate cages at Avian Conservation and Research Center, Department of Wildlife an Ecology, University of Veterinary and Animal Sciences, Lahore, Pakistan. 100 fecal and 100 blood samples for each bird species were inspected to analyze internal parasites. During present study, 17 species of endoparasites 14 from fecal samples and three from blood were examined. Two species of ectoparasites i.e. mite Dermanyssus gallinae 42% and fowl ticks Args persicus 41%were studied. Blood parasites included Plasmodium juxtanucleare 50%, Leucoctoyzoon simond having parasitic prevalence 40%, and Aegyptinella pullorum having parasitic prevalence of 40%. Parasitic species recorded from fecal samples included 6 species of nematodes viz. Allodpa suctoria 2%. Syngamus trachea with parasitic prevalence of 60%, Capillaria annulata 37.5%, Ascardia galli 24%, Capillaria anatis 40% and Heterakis gallinarum 28.3%. Similarly, two species of trematodes viz. Prosthogonimus ovatus having parasitic prevalence of 50% and Prosthogonimus macrorchis 21% were also documented from fecal avian samples . Single cestode species Raillietina echinobothrida having parasitic prevalence of 72% and 3 protozoan species i.e. Eimeria maxima having parasitic prevalence of 21%, Giardia lamblia 41% and Histomonas meleagridis 18% were documented during corpological analysis. In our recommendation, proper sanitation, medication and vaccination of bird’s enclousres are suggested to avoid parasites. RESUMO Amostras de sangue e fezes de perdiz chukar (Alectoris chukar), faisão-albino (Phasianus colchicus), faisão-prateado (Lophura nycthemera), periquito-de-rosa (Psittacula krameri) e perus (Meleagris gallopavo) foram analisadas para verificar a prevalência de parasitas. Para registrar os parasitas, essas cinco espécies de aves foram colocadas em gaiolas separadas no Centro de Conservação e Pesquisa de Aves, Departamento de Vida Selvagem e Ecologia, Universidade de Veterinária e Ciências Animais, Lahore, Paquistão. Cem amostras fecais e 100 amostras de sangue para cada espécie de ave foram inspecionadas para analisar os parasitas internos. Durante o presente estudo, foram examinadas 17 espécies de endoparasitas, 14 de amostras fecais e 3 de sangue. Foram estudadas duas espécies de ectoparasitas, ou seja, o ácaro Dermanyssus gallinae 42% e o carrapato aviário Args persicus 41%. Os parasitas sanguíneos incluíram Plasmodium juxtanucleare 50%, Leucoctoyzoon simond com prevalência parasitária de 40% e Aegyptinella pullorum com prevalência parasitária de 40%. As espécies parasitas registradas em amostras fecais incluíram 6 espécies de nematoides viz. Allodpa suctoria 2%, Syngamus traqueia com prevalência parasitária de 60%, Capillaria annulata 37,5%, Ascardia galli 24%, Capillaria anatis 40% e Heterakis gallinarum 28,3%. Da mesma forma, duas espécies de trematódeos viz. Prosthogonimus ovatus com prevalência parasitária de 50% e Prosthogonimus macrorchis 21% também foram documentados em amostras fecais de aves. Espécies de cestoide único Raillietina echinobothrida com prevalência parasitária de 72% e 3 espécies de protozoários, isto é, Eimeria maxima com prevalência parasitária de 21%, Giardia lamblia 41% e Histomonas meleagridis 18% foram documentadas durante a análise corpológica. Em nossa recomendação, o saneamento adequado, medicação e vacinação de invólucros de pássaros são sugeridos para evitar parasitas.

During this one year study, blood and fecal samples of doves (Zenaida asiatica), ducks (Anas platyrhynchos), pigeons (Columba livia), partridges (Alectoris chukar), turkeys (Meleagris gallopavo) and goose (Chen caerulescens) were collected to assess the parasitic prevalence in these birds. The birds were kept at Avian Conservation and Research Center, Department of Wildlife and Ecology, University of Veterinary and Animal Sciences, Lahore. All these avian species were kept in separate cages and their entire body was inspected on regularly basis to record external parasites. For internal parasites, 100 blood and 100 fecal samples for each species were analyzed. During present study, two species of ectoparasites i.e. fowl ticks (Args persicus) and mite (Dermanyssus gallinae) while 17 species of endoparasites; three from blood and 14 from fecal samples were identified. Prevalence of blood parasites was Plasmodium juxtanucleare 29.3%, Aegyptinella pullorum 15% and Leucoctoyzoon simond 13%. Parasitic species recorded from fecal samples included 6 species of nematodes viz. Syngamus trachea with parasitic prevalence of 50%, Capillaria anatis 40%, Capillaria annulata 37.5%, Heterakis gallinarum 28.3%, Ascardia galli 24% and Allodpa suctoria 2%. Similarly, two species of trematodes viz. Prosthogonimus ovatus having parasitic prevalence of 12.1% and Prosthogonimus macrorchis 9.1% were also recorded from fecal samples of the birds. Single cestode species Raillietina echinobothrida having parasitic prevalence of 27% and 3 protozoan species i.e. Eimeria maxima having prevalence 20.1%, Histomonas meleagridis 8% and Giardia lamblia 5.3% were recorded. In our recommendation, proper medication and sanitation of the bird’s houses and cages is recommended to avoid parasites. Resumo Durante este estudo de um ano, amostras de sangue e fezes de pombos (Zenaida asiatica), patos (Anas platyrhynchos), pombos (Columba livia), perdizes (Alectoris chukar), perus (Meleagris gallopavo) e ganso (Chen caerulescens) foram coletados para avaliar a prevalência de parasitas nessas aves. As aves foram mantidas no Centro de Conservação e Pesquisa de Aves, Departamento de Vida Selvagem e Ecologia, Universidade de Veterinária e Ciências Animais, Lahore. Todas essas espécies de aves foram mantidas em gaiolas separadas e todo o seu corpo foi inspecionado regularmente para registrar parasitas externos. Para parasitas internos, foram analisadas 100 amostras de sangue e 100 amostras fecais de cada espécie. Durante o presente estudo, duas espécies de ectoparasitas, ou seja, carrapatos de aves (Args persicus) e ácaros (Dermanyssus gallinae), enquanto 17 espécies de endoparasitas, três de sangue e 14 de amostras fecais, foram identificadas. Os parasitas sanguíneos prevalentes foram Plasmodium juxtanucleare, 29,3%, Aegyptinella pullorum, 15%, e Leucoctoyzoon simond, 13%. As espécies parasitas registradas em amostras fecais incluíram 6 espécies de nematoides viz. Syngamus traqueia com prevalência parasitária de 50%, Capillaria anatis, 40%, Capillaria annulata, 37,5%, Heterakis gallinarum, 28,3%, Ascardia galli, 24% e Allodpa suctoria, 2%. Da mesma forma, duas espécies de trematódeos viz. Prosthogonimus ovatus com prevalência parasitária de 12,1% e Prosthogonimus macrorchis, 9,1%, também foram registrados nas amostras fecais das aves. Espécies de cestoide único Raillietina echinobothrida com prevalência parasitária de 27% e 3 espécies de protozoários, ou seja, Eimeria maxima tendo prevalência de 20,1%, Histomonas meleagridis, 8%, e Giardia lamblia, 5,3%, foram registradas. Em nossa recomendação, são indicados medicação adequada e saneamento das casas e gaiolas dos pássaros para evitar parasitas.

Leucocytozoon has been globally described as avian hosts, and it significantly affects many avian taxa including the fighting cock. The clinical signs of leucocytozoonosis range from asymptomatic to high morbidity leading to increase mortality rates. Interestingly, there are insufficient epidemiological studies of this blood parasite or the molecular identification of infections within the vertebrate host. In this study, 250 blood samples were collected from fighting cocks at 9 districts in Maha Sarakham Province, Thailand. Leucocytozoon infections were screened by blood examination and nested PCR followed by sequence analysis of mitochondrial cytochrome b gene was used to identify Leucocytozoon species. Twenty-two out of 250 (8.8%) samples had confirmed Leucocytozoon infections based on microscopic examination whereas with nested-PCR, 52 samples tested positive. Of these 52, 51 were successfully sequenced among which, one was Plasmodium juxtanucleare, 45 were Leucocytozoon sp. (18%) and 5 were L. schoutedeni (2%). This study is the first report to demonstrate the molecular prevalence of leucocytozoonosis in fighting cock in Thailand. This study indicated that leucocytozoonosis is widespread in fighting cock although the frequency was not determined and needs further study.

Culicoides biting midges are vectors of many haemosporidian parasite species. These parasites are found in several wild and domestic avian species in Thailand but knowledge of the insect vectors is very limited. In this study, a molecular approach was used to detect haemosporidian parasites in six biting midge species in Thailand. A total of fifteen cytochrome b haplotypes of three haemosporidian parasites were detected from 1,165 specimens of six different Culicoides species. Comparisons of these sequences with those recorded in a public database revealed that they were comprised sequences of three species, Leucocytozoon sp., Plasmodium juxtanucleare and P. gallinaceum. All of these haemosporidian parasite species were detected in Culicoides mahasarakhamense Pramual, Jomkumsing, Piraonapicha, & Jumpato while P. juxtanucleare was also detected in C. huffi Causey and C. guttifer Meijere and Leucocytozoon sp. were also detected in C. guttifer. All of these parasites are commonly found in chickens in agreement with information that these biting midge species will bite chickens. Detection DNA of Leucocytozoon sp. in biting midges reported here provides the second record of this parasitic genus infecting Culicoides. This study also provides the first records of P. juxtanucleare and P. gallinaceum in biting midge species. Further investigation is required to determine whether Culicoides biting midge species are competent vectors of these parasites.

Haemosporidian infections in domestic chickens (Gallus gallus domesticus) are not only widely prevalent but also cause economic loss. Diagnosis is usually made by microscopic examination; however, the method has several drawbacks such as requiring an experienced microscopist, being unreliable when parasitemia is low and being unable to accurately differentiate between co-infections from multiple parasite species. Therefore, the current extent of haemosporidian infections might be underestimated and neglected. We have developed a novel multiplex PCR assay to simultaneously detect and differentiate between four haemosporidian parasites: Leucocytozoon caulleryi, Leucocytozoon sabrazesi, Plasmodium juxtanucleare and Plasmodium gallinaceum. Primers in the present study specifically amplified the corresponding targets with no cross-species amplification detected. The multiplex PCR exhibited a significantly greater detection rate when compared with microscopic examination (p = 0.0001). The results demonstrate that the detection rate of multiplex PCR for L. sabrazesi, P. juxtanucleare, and P. gallinaceum are all greater than that of microscopic examination with p = 0.002, 0.0001 and 0.004, respectively. Co-infections were also detected more effectively by multiplex PCR. We applied the current method to field samples originating from Nan, Prachinburi, and Chachoengsao Provinces. The current study revealed that positive rates of haemosporidian parasites in chickens in the three study sites ranging from 39.5%–93.8%. The present assay offers a timesaving option for molecular diagnosis instead of using singleplex PCRs for detecting the parasites individually. Within a single reaction, this assay would be a useful tool for the detection of avian haemosporidian parasites either single or under co-infection conditions and for large-scale epidemiology studies.

application/pdf, Plasmodium gallinaceum and Plasmodium juxtanucleare are the most common species of Plasmodium causing avian malaria in domestic and wild chickens. In Thailand, P. gallinaceum infection in poultry has been reported since 1996, but P. juxtanucleare infection has never been reported. The objective of this study was to identify and characterize the Plasmodium species infecting Burmese red junglefowls (Gallus gallus spadiceus) in Thailand using molecular techniques. Seventeen Burmese red junglefowls from a farm in western Thailand were suspected of being infected with blood parasite. Blood samples of all seventeen Burmese red junglefowls were collected and examined. Microscopic examination revealed the presence of Plasmodium trophozoites in 5 out of 17 samples. In comparison, nested PCR detected 12 samples to be positive for Plasmodium spp. Molecular characterization, based on partial sequence of mitochondrial cytochrome b gene, revealed 7 PCR-positive samples as P. juxtanucleare. Nucleotide sequence comparison and phylogenetic analyses grouped these 7 P. juxtanucleare into 2 distinct groups. The first group was identical to P. juxtanucleare previously reported from Brazil, Malaysia and Taiwan, whereas the second group was identical to P. juxtanucleare from Japan. This is the first report on molecular confirmation and characterization of P. juxtanucleare in Burmese red junglefowls in Thailand.

Village chicken production, a traditional, small-scale, and extensive backyard poultry industry, has been profitable for local farmers in Myanmar. However, there is scanty information available concerning the infection of these chickens with avian pathogens, including haemoprotozoan parasites. In the present study, we provide the first report of microscopic detection and molecular identification of Leucocytozoon and Plasmodium parasites from seven different areas of Myanmar. Leucocytozoon gametocytes were detected in 17.6% (81/461) of the blood smears from village chickens. The nested polymerase chain reaction (PCR) for targeting Leucocytozoon mitochondrial cytochrome b (cyt b) genes had a 17.6% positive rate. Although the positive rate of nested PCR targeting Plasmodium/Haemoproteus cyt b was 34.3%, the PCR protocol was observed to possibly amplify DNA of a certain species of Leucocytozoon. There were no obvious clinical signs in the infected birds. Statistical analysis of the microscopic detection and PCR detection rates using the age and sex of birds as internal factors revealed that the statistical significances differed according to the study area. The sequencing of 32 PCR products obtained from each study area revealed infection by Leucocytozoon caulleryi in three birds, Leucocytozoon sabrazesi in two birds, Leucocytozoon schoutedeni in two birds, Leucocytozoon sp. in eighteen birds, and Plasmodium juxtanucleare in seven birds; however, Haemoproteus infection was not detected. While L. sabrazesi was detected in chickens from the central region of Myanmar, the other haemosporidians were detected in those from different areas. In the haplotype analysis, we detected 17 haemosporidian cyt b haplotypes, including two for L. caulleryi, one for L. sabrazesi, two for L. schoutedeni, nine for Leucocytozoon sp., and three for P. juxtanucleare. Phylogenetic analysis of the cyt b haplotypes revealed a considerably close genetic relationship among chicken haemosporidians detected in Myanmar, Thailand, and Malaysia. These results indicate that well-recognized widespread species of chicken Leucocytozoon and Plasmodium are distributed nationwide in Myanmar, providing new insights into the ecosystem and control strategies of haemosporidian parasites in domesticated chickens in Myanmar.

SUMMARYThe pathogensPlasmodium juxtanucleareand chicken anaemia virus (CAV) are easily transmitted and potentially harmful to chickens. In this study, we established an experimental model to investigate the effects of avian malaria caused byP. juxtanuclearein white leghorn specific-pathogen-free (SPF) chicks previously immunosuppressed with CAV. Parasitaemia, haematological variables and clinical and pathological parameters were determined in four different experimental groups: chicks coinfected by CAV andP. juxtanuclearestrain (Coinfected group), chicks exclusively infected by CAV (CAV group) orP. juxtanucleare(Malaria group) and uninfected chicks (Control group). Our data demonstrated thatP. juxtanucleareparasitaemia was significantly higher in the Coinfected group. Furthermore, haematological parameters, including the RBC, haematocrit and haemoglobin concentration were significantly reduced in coinfected chicks. In agreement with the changes observed in haematological features, the mortality among coinfected chicks was higher compared with animals with single infections. Clinical analysis indicated moderate changes related to different organs size (bursa of Fabricius, heart and liver) in coinfected birds. The experimental coinfection of SPF chickens withP. juxtanucleareand CAV may represent a research tool for the study of avian malaria after CAV immunosuppression, enabling measurement of the impacts caused by different pathogens during malarial infection.

One of the species that causes avian malaria is Plasmodium juxtanucleare. It is commonly found in poultry, especially when the birds receive food free of coccidiostats. Since industrial and organic poultry breeding is increasing in the world and few studies have been conducted examining the clinical parameters of both healthy and infected birds, this work evaluated whether the infection caused by P. juxtanucleare in Gallus gallus provokes alterations in the birds’ hepatic profile. We analyzed the activity of ALT and AST and carried out histological analyses of liver sections of infected fowls by intracelomic inoculation with infected blood from a donor fowl with a parasite load of around 7%. The infected birds’ parasite load was evaluated during 45 days by means of blood smears. There was a positive correlation between the increase in parasite load and higher ALT activity in the infected fowls, but there was no significant variation of the AST activity between the control and infected groups, possibly because of the non-specificity of this enzyme as an indicator of hepatic lesion. The results show that infection caused by P. juxtanucleare in G. gallus provokes hepatic alterations, indicated by the increase in the ALT enzyme activity and by the inflammatory infiltrates found in the liver sections of the infected fowls.

Background: The Thailand-Myanmar borderland is an area endemic for malaria where transmission is low, seasonal and unstable. The epidemiology has been described but there is relatively few data on the entomological determinants of malaria transmission. Methods: Entomological investigations were conducted during 24 months in four villages located in Kayin state, on the Myanmar side of the Thailand-Myanmar border. Anopheles mosquitoes were identified by morphology, and molecular assays were used in order to discriminate between closely related sibling species of malaria vectors. Plasmodium infection rate was determined using quantitative real-time PCR. Results: The diversity of Anopheles mosquitoes was very high and multiple species were identified as malaria vectors. The intensity of human-vector contact (mean human-biting rate= 369 bites/person/month) compensates for the low infection rate in naturally infected populations of malaria vectors (mean sporozoite index= 0.04 and 0.17 % for P. falciparum and P. vivax respectively), yielding intermediary level of transmission intensity (mean entomological inoculation rate= 0.13 and 0.64 infective bites/person/month for P. falciparum and P. vivax, respectively). Only 36% of the infected mosquitoes were collected indoors between 09:00 pm and 05:00 am, suggesting that mosquito bed-nets would fail to prevent most of the infective bites in the study area. Conclusion: This study provided a unique opportunity to describe the entomology of malaria in low transmission settings of Southeast Asia. Our data are important in the context of malaria elimination in the Greater Mekong Subregion.

ELISEI C.; FERNANDES, K.R.; FORLANO, M.D.; MADUREIRA, R.C.; SCOFIELD, A.; YOTOKO, K.C.; SOARES C.O.; ARAUJO, F.R.; MASSARD C.L. Morphology and morphometry of three Plasmodium juxtanucleare (Apicomplexa: Plasmodiidae) isolates. (Morfologia e morfometria de tres isolados de Plasmodium juxtanucleare (Apicomplexa: Plasmodiidae)). Revista Brasileira de Parasitologia Veterinaria, v. 16, n. 3, p. 139-144, 2007. Laboratorio de Biologia Molecular Sanidade Animal, Embrapa Gado de Corte, Campo Grande, MS, Brasil. E-mail: carinaelisei@yahoo.com.br In this work, three isolates of Plasmodium juxtanucleare have been analyzed based on morphological, morphometric and parasitic parameters. Each isolate was sampled from naturally infected adult chicken (Gallus gallus) from rural areas of three Brazilian municipalities: Seropedica (22° 48' S; 43° 41' W), in the state of Rio de Janeiro; Cruzeiro (22° 33' S; 44° 57' W), in the state of Sao Paulo; and Santa Barbara do Tugurio (21° 15' S; 43° 27' W), in the state of Minas Gerais. The blood samples taken from each infected chicken were inoculated in three groups of ten young chicken (21 days old). Blood smears of the experimentally infected chicken were sampled every two days until the 69th day in order to evaluate the parasitemia. For the morphological-descriptive and morphometric analyses, we measured 30 individuals from each of the intraerythocytic states, measures of the major (MD) and minor diameters (md), the estimation of morphometric index (Mi=md/MD) and size (T=pab, a= md/2; b=MD/2). The results indicated low and homogeneous parasitemia rates in the three strains, which showed differences among shape and size of the parasitic stadia displayed.

Submitted by Jorge Silva (jorgelmsilva@ufrrj.br) on 2017-06-26T17:32:22Z No. of bitstreams: 1 2015 - Thamyris Sampaio Vilela.pdf: 3001387 bytes, checksum: d945149445f8ee2765222f5475c7eaa2 (MD5) Made available in DSpace on 2017-06-26T17:32:22Z (GMT). No. of bitstreams: 1 2015 - Thamyris Sampaio Vilela.pdf: 3001387 bytes, checksum: d945149445f8ee2765222f5475c7eaa2 (MD5) Previous issue date: 2015-02-27 CAPES - Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior CNPq - Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico FAPERJ - Funda??o Carlos Chagas Filho de Amparo ? Pesquisa do Estado do Rio de Janeiro This study aimed to develop a real-time PCR (qPCR) assay for diagnosis of Plasmodium spp. using as target the 18S rDNA and Cyt b genes. A range of 101 blood samples were collected from Gallus gallus in poultry rustic breeds of in Rio de Janeiro, Brazil. The collected bloods were used to prepare blood smears and to extract the deoxyribonucleic acid (DNA) of these samples. There with, molecular protocols were tested, such as the already published conventional PCR (cPCR) and nested PCR (nPCR), designed for Plasmodium spp. In this study two qPCR protocols were developed using primers targeting the Cyt b and 18S rDNA genes. The qPCR detection limit for both genes was 10 copies of the target DNA, which were higher than the detection limit observed in nPCR and cPCR. In qPCR, 69.30% (n = 70/101) samples were positive targeting 18Sr DNA gene and 59.40% (n = 60/101) samples were positive targeting Cyt b gene. In nPCR and cPCR, 54.45% (n = 55/101) and 52.47% (n= 53/101) samples were positive, respectively. In blood smear microscopy, 31 (30.69%) samples were positive. There was no disagreement between the results (p > 0.05) of qPCR for 18Sr DNA and Cyt b genes. Additionally, qPCR was more sensitive than the other techniques discussed, mostly related to blood smear microscopy (p < 0.05). Therefore, the two qPCR developed in the study showed more sensitivity than other techniques and enabled the detection of Plasmodium spp. in poultry even in low parasitemia. Este estudo teve como objetivo desenvolver um ensaio de PCR em tempo real (qPCR) para o diagn?stico de Plasmodium spp. utilizando os genes 18S rDNA e cyt b. Foram coletadas 101 amostras de sangue de aves da esp?cie Gallus gallus de cria??o r?stica ou org?nica no munic?pio de Serop?dica, Rio de Janeiro. Os sangues coletados foram utilizados na prepara??o de esfrega?os de sangue e para extra??o do ?cido desoxirribonucl?ico (DNA) destas amostras. Protocolos de ensaios moleculares foram testados, tal como o PCR convencional (cPCR), nestedPCR (nPCR) e qPCR para Plasmodium spp. Neste estudo dois protocolos de qPCR foram desenvolvidos utilizando oligoiniciadores desenhados com alvo no citocromo b e 18S rDNA. O limite de detec??o para os dois genes qPCR foi de 10 c?pias do alvo de DNA, que foram maiores do que o limite de detec??o observado em nPCR e cPCR. Em qPCR, 69,30% (n = 70/101) amostras foram positivas com alvo no gene 18SrDNA e 59,40%(n = 60/101) amostras positivas com alvo no gene cyt b. Em nPCR e cPCR, 54,45% (n = 55/101) e 52,47% (n = 53/101) amostras foram positivas, respectivamente. Em microscopia de esfrega?o de sangue, 31 (30,69%) amostras foram positivas. N?o houve discord?ncia entre os resultados (p> 0,05) de qPCR para os genes 18SrDNA e cyt b.. Al?m disso, qPCR foi mais sens?vel do que as outras t?cnicas discutidas, principalmente relacionado com a microscopia ?ptica (p

Plasmodium juxtanucleare, uma de duas espécies causadoras de malária em galinhas, pode ser encontrada em diferentes países do mundo, incluindo o Brasil. O objetivo geral deste estudo foi avaliar a infecção por P. juxtanucleare em pintinhos livres de patógenos (SPF) e previamente imunossuprimidos pelo vírus da anemia infecciosa das galinhas (CAV). Para o estabelecimento deste modelo foram abordadas duas estratégias experimentais: (i) avaliação de alterações hematológicas, biológicas e patológicas, (ii) avaliação da resposta celular das aves infectadas. Para isso, definiram-se quatro grupos experimentais: Grupo P. juxtanucleare: aves infectadas exclusivamente por P. juxtanucleare, Grupo CAV: aves infectadas exclusivamente por CAV, Grupo co-infectado: aves co-infectadas por P. juxtanucleare e por CAV e Grupo controle: aves não infectadas. A parasitemia por P. juxtanucleare apresentou-se baixa em ambos os grupos infectados por P. juxtanucleare (grupos P. juxtanucleare e co-infectado); entretanto, valores significativamente maiores foram detectados nas aves co-infectadas. Anemia foi observada nos três grupos infectados. No entanto, aves co-infectadas apresentaram anemia mais severa, observada pela redução significativa do hematócrito quando comparadas aos outros grupos infectados. A mortalidade foi maior e detectada precocemente no grupo co-infectado, entretanto as lesões patológicas foram moderadas quando comparadas às lesões acentuadas observadas nas aves dos grupos P. juxtanucleare e CAV. Esses resultados indicam que os microrganismos apresentam efeito patogênico potencializado durante ambas às infecções. O número total de leucócitos e de linfócitos apresentaram redução significativa (p

This study aimed to evaluate the 4-(6′-thiopurine)-7-chloroquinoline, a novel quinoline/6-thiopurine conjugate, for the treatment of Gallus gallus experimentally infected with Plasmodium juxtanucleare, an avian malaria agent. The avian group treated with 4-(6′-thiopurine)-7-chloroquinoline showed a significative parasite clearance and maintained a low level of parasitaemia, when compared with the untreated control group and to the chloroquine treated avian group.

Plasmodium juxtanucleare is one of the agents that cause chicken malaria. High parasitemia causes anemia, diarrhea and weight loss that leads to death. This parasite is spread through the tropics and little is known about the physiological changes caused by P. juxtanucleare infections in chickens. The aim of this work was to determine hematological changes in chickens experimentally infected with this parasite. No weight or temperature differences between infected and control groups were detected. Low parasitemia was observed reaching a peak after 15 days; trophozoits was the most observed form, followed by schizonts detected on day 12 and gametocytes on day 27. Infected thrombocytes and thrombocytes with pyknotic nuclei and retracted cytoplasm were observed along the experimental infection. No changes were observed on the erythrocyte values. However, a reduction of thrombocyte number and an increase of total leukocyte and basophil numbers, all significant, were detected in infected chickens. Parasitemia was probably kept low by general immunological activation. These results suggest that classical hematology can monitor physiological changes caused by P. juxtanucleare infections in chickens.

Plasmodium juxtanucleare is one of the agents that cause avian malaria in chickens and little is known about the treatment of the infection. The aim of this study was to compare the antimalarial activities of rutin and chloroquine in chickens infected with P. juxtanucleare after immunossuppression with corticosteroid. Antimalarial activity was verified in 33 fowls by evaluation of parasitemia levels in Giemsa-stained blood smears taken to 30 days. Rutin did not exert a reduction in parasitemia from P. juxtanucleare when compared to chloroquine treatment that kept the parasites at a low level, demonstrating its antimalarial effect. During the course of the infection, the trophozoite stage predominated (80%), followed by schizont (17%) and gametocyte (3%). Maximum parasitemia levels were recorded on day 12 in control and rutin groups. There were no significant differences in the hematocrit values, weight body or the temperature of the fowls among the groups evaluated (p > 0.05).

In this work, three isolates of Plasmodium juxtanucleare have been analyzed based on morphological, morphometric and parasitic parameters. Each isolate was sampled from naturally infected adult chicken (Gallus gallus) from rural areas of three Brazilian municipalities: Seropédica (22º 48' S; 43º 41'W), in the state of Rio de Janeiro; Cruzeiro (22º 33' S; 44º 57'W), in the state of São Paulo; and Santa Bárbara do Tugúrio (21º 15' S; 43º 27' W), in the state of Minas Gerais. The blood samples taken from each infected chicken were inoculated in three groups of ten young chicken (21 days old). Blood smears of the experimentally infected chicken were sampled every two days until the 69th day in order to evaluate the parasitemia. For the morphological-descriptive and morphometric analyses, we measured 30 individuals from each of the intraerythocytic states, measures of the major (MD) and minor diameters (md), the estimation of morphometric index (Mi=md/MD) and size (T=pab, a= md/2; b=MD/2). The results indicated low and homogeneous parasitemia rates in the three strains, which showed differences among shape and size of the parasitic stadia displayed. Neste trabalho, três isolados de Plasmodium juxtanucleare foram analisados com base na morfologia, morfometria e parâmetros parasitológicos. Cada isolado foi coletado de aves (Gallus gallus) adultas infectadas naturalmente de áreas rurais de três municípios brasileiros: Seropédica (22º 48' S; 43º 41' W), no estado do Rio de Janeiro; Cruzeiro (22º 33' S; 44º 57' W), no estado de São Paulo; e Santa Bárbara do Tugúrio (21º 15' S; 43º 27' W), no estado de Minas Gerais. As amostras de sangue coletadas de cada ave infectada foram inoculadas em três grupos de dez aves (21 dias de idade). Esfregaços sangüíneos das aves infectadas experimentalmente foram realizados de dois em dois dias durante um período de 69 dias para avaliar a parasitemia. Para análises morfofisiológica e morfométrica, foram mensurados 30 indivíduos de cada estágio intraeritrocítico. Foram tomadas as medidas do diâmetro maior (DM) e diâmetro menor (dm), com os quais foi estimado o índice morfométrico (Mi=md/MD) e o tamanho (T=pab, a= md/ 2; b=MD/2). Os resultados indicaram uma parasitemia homogênea entre os três isolados, havendo diferenças apenas nas formas e tamanhos dos estádios parasitários.

Neste trabalho, três isolados de Plasmodium juxtanucleare foram analisados com base na morfologia, morfometria e parâmetros parasitológicos. Cada isolado foi coletado de aves (Gallus gallus) adultas infectadas naturalmente de áreas rurais de três municípios brasileiros: Seropédica (22º 48' S; 43º 41' W), no estado do Rio de Janeiro; Cruzeiro (22º 33' S; 44º 57' W), no estado de São Paulo; e Santa Bárbara do Tugúrio (21º 15' S; 43º 27' W), no estado de Minas Gerais. As amostras de sangue coletadas de cada ave infectada foram inoculadas em três grupos de dez aves (21 dias de idade). Esfregaços sangüíneos das aves infectadas experimentalmente foram realizados de dois em dois dias durante um período de 69 dias para avaliar a parasitemia. Para análises morfofisiológica e morfométrica, foram mensurados 30 indivíduos de cada estágio intraeritrocítico. Foram tomadas as medidas do diâmetro maior (DM) e diâmetro menor (dm), com os quais foi estimado o índice morfométrico (Mi=md/MD) e o tamanho (T=pab, a= md/ 2; b=MD/2). Os resultados indicaram uma parasitemia homogênea entre os três isolados, havendo diferenças apenas nas formas e tamanhos dos estádios parasitários. In this work, three isolates of Plasmodium juxtanucleare have been analyzed based on morphological, morphometric and parasitic parameters. Each isolate was sampled from naturally infected adult chicken (Gallus gallus) from rural areas of three Brazilian municipalities: Seropédica (22º 48' S; 43º 41'W), in the state of Rio de Janeiro; Cruzeiro (22º 33' S; 44º 57'W), in the state of São Paulo; and Santa Bárbara do Tugúrio (21º 15' S; 43º 27' W), in the state of Minas Gerais. The blood samples taken from each infected chicken were inoculated in three groups of ten young chicken (21 days old). Blood smears of the experimentally infected chicken were sampled every two days until the 69th day in order to evaluate the parasitemia. For the morphological-descriptive and morphometric analyses, we measured 30 individuals from each of the intraerythocytic states, measures of the major (MD) and minor diameters (md), the estimation of morphometric index (Mi=md/MD) and size (T=pab, a= md/2; b=MD/2). The results indicated low and homogeneous parasitemia rates in the three strains, which showed differences among shape and size of the parasitic stadia displayed.

We analyzed mitochondrial genomes of two avian malaria protozoa, Plasmodium gallinaceum and Plasmodium juxtanucleare. Both mitochondrial genomes were estimated to be 6,002 and 6,014 bp in length, respectively, and to have the identical gene organization and contents to that of other Plasmodium species previously analyzed; three functional genes for cytochrome c oxidase subunit I, III, and cytochrome (cyt b), with following sets of discontinuous and scrambled 15 ribosomal subunit RNA (rRNA) genes. Similarities of the three protein-coding genes showed closer relationship within avian malaria protozoa rather than mammalian Plasmodium species. In addition, we showed the tandem repeated structure of each mitochondrial genome of both P. gallinaceum and P. juxtanucleare as well as previously found in mammalian Plasmodium species. This study revealed the complete sequences and structure of the mitochondrial genomes of avian malaria protozoa for the first time.

Este trabalho sobre a malária aviária causada pelo Plasmodium juxtanucleare em Gallus gallus no Brasil, foi realizado no sítio Boa Vista (21º15'12"S, 43º27'16"W), município de Santa Bárbara do Tugúrio, Minas Gerais, e teve como objetivos reavaliar a prevalência de 100% encontrada em estudos anteriores e acompanhar a variação dos estádios eritrocíticos do parasito ao longo de um ano na região. Sob microscopia em imersão foram examinados esfregaços sangüíneos de 25 aves mestiças, corados pelo Giemsa. As amostras foram coletadas ao longo de um ano, mensalmente de novembro/00 a maio/01 e quinzenalmente de junho/01 a outubro/01. Os estádios eritrocíticos foram quantificados pela observação de 100 campos microscópicos. Foi constatada prevalência de 100% por P. juxtanuclerare na região. Não houve correlação estatisticamente significativa entre o aumento dos estádios eritrocíticos em relação às coletas ao longo do ano. O estádio de trofozoíto foi predominante durante todo o período de estudo The avian malaria caused by Plasmodium juxtanucleare in Gallus gallus, is a tipical plasmodiose from Brazilian gallinaceous. This disease can causes morbidy and mortality in its vertebrate hosts. This work was conducted at Boa Vista farm, Municipality of Santa Bárbara do Tugúrio, Minas Gerais, Brazil and the ours objectives were to evaluate the hight prevalence found in previous studies and to accompany the variation of the erytrocytic forms during one year. The bloods smears, dyed with Giemsa were examined in microscopy immersion. Twenty five half-breed fowls were accompany duting one year, monthly (from November/00 to May/01) and biweekly (from June/01 to October/01). The erytrocytic forms were registered and quantified by the observation of 100 microscopic fields. Was verified a prevalence of 100% by P. juxtanuclerare, but there wasn't statistics correlation between the increase of the erytrocytic forms during the year. The trophozoites were the more abundant form found in this studie

The avian malaria caused by Plasmodium juxtanucleare in Gallus gallus, is a tipical plasmodiose from Brazilian gallinaceous. This disease can causes morbidy and mortality in its vertebrate hosts. This work was conducted at Boa Vista farm, Municipality of Santa Barbara do Tugurio, Minas Gerais, Brazil and the ours objectives were to evaluate the hight prevalence found in previous studies and to accompany the variation of the erytrocytic forms during one year. The bloods smears, dyed with Giemsa were examined in microscopy immersion. Twenty five half-breed fowls were accompany duting one year, monthly (from November/00 to May/01) and biweekly (from June/01 to October/01). The erytrocytic forms were registered and quantified by the observation of 100 microscopic fields. Was verified a prevalence of 100% by P. juxtanuclerare, but there wasn’t statistics correlation between the increase of the erytrocytic forms during the year. The trophozoites were the more abundant form found in this studie.

Five black-footed penguins (Spheniscus demersus) admitted to the Southern African Foundation for the Conservation of Coastal Birds, in Cape Town, South Africa, died from malaria infection. Evidence for malaria as the cause of death included antemortem clinical signs, parasitemia, splenomegaly, pulmonary edema, and the presence of histologically visible schizonts in the reticuloendothelial system. A portion of the malarial small subunit ribosomal ribonucleic acid gene was detected by polymerase chain reaction from postmortem blood samples from all the birds. A species-specific variable region of this gene was compared with the same region on genes from other known avian malarial organisms, establishing that Plasmodium juxtanucleare was involved.

International audience; Plasmodium (Novyella) juxtanucleare is a widely distributed parasite that primarily infects chickens (Gallus gallus domesticus). All species of Novyella are characterized by very small schizonts, which in the case of P. juxtanucleare are always found juxtaposed to the erythrocyte nucleus, hence its name. Nearly complete small-subunit ribosomal RNA sequences have been obtained from 2 isolates of this species, and comparisons with other Plasmodium species have been made. Phylogenetic analysis reveals that this parasite is closely related to other avian-infecting Plasmodium species and that molecular relationships among the avian-infecting plasmodia do not correspond to their morphology-based subgeneric classifications.

Se realizó el análisis morfométrico y morfológico de un aislado de (MSS-47) de Plasmodium juxtanucleare. Fueron evaluados, la parasitemia, las medidas del diámetro menor (dm), el diámetro mayor (DM) y el índice morfométrico (IM = dm/DM) de formas intraeritrociticas, aves inoculadas experimentalmente. En 106 trofozoítos se observaron formas puntiformes, ovoides, anilladas, elípticas, piriformes y redondeadas, con predominio de las formas ovoides y elípticas. La media de los trofozoítos fue de 1.802 ± 0,313 x 1.230 ± 0,2 mm, con un IM = 0,623 ± 0,110 mm. En 98 esquizontes, se observaron formas ovoides, elípticas piriformes y ameboides, con predominio de formas ovoides y piriformes. La media de los esquzontes fue de 2.809 ± 0,187 x 1.820 ± 0,386 mm, con un IM = 0,643 ± 0,120. En 18 macrogametocitos, se observaron formas ovoides, redondeadas, elípticas y piriformes, con predominio de las formas ovoides y elípticas. La media de los macrogametocitos fue de 3.559 ± 0,883 x 2.363 ± 0,207 mm, con un IM = 0,766 ± 0,078 mm. En 34 microgametocitos, se observaron formas ovoides, redondeadas, elípticas y piriformes, con predominio de las formas ovoides y elípticas. La media de los microgametocitos fue de 3.891 ± 0,946 x 2.386 ± 0,783 mm, con un IM = 0,505 ± 0,103 mm. Por lo tanto, el aislado MSS-47 de P. juxtanucleare se puede caracterizar como muy pleomórfico The morphological and morphometric analysis of Plasmodium juxtanucleare (MSS-47) isolate were done. The parasitemia was evaluated, and the smaller diameter (sd), the bigger diameter (BD) and the morphometric index (MI = sd/BD) of erythrocytic forms from experimentally inoculated chickens, were measured. In 106 trophozoites, were showed point, ovoid, ring, elliptical, pear and round forms, with predominance of the ovoid and elliptical forms. The average measure of the trophozoites was 1.802 ± 0.313 x 1.230 ± 0.208 mm, with MI = 0.623 ± 0.110 mm. In 98 schizonts, were showed ovoid, elliptical, pear and ameboid forms, with predominance of the ovoid and pear forms. The average measure of the schizonts was 2.809 ± 0.187 x 1.820 ± 0.386 mm, with MI = 0.643 ± 0.120 mm. In 18 macrogametocytes, were showed ovoid, round, elliptical and pear forms, with predominance of the ovoid and elliptical forms. The average measure of the macrogametocytes was 3.559 ± 0.883 x 2.363 ± 0.207 mm, with MI = 0.766 ± 0.078. In 34 microgametocytes, were showed ovoid, round, elliptical and pear forms, with predominance of the ovoid and elliptical forms. The average measure of the microgametocytes was 3.891 ± 0.946 x 2.386 ± 0.783 mm, with MI = 0.505 ± 0.103. The MSS-47 isolate of P. juxtanucleare was characterized as highly pleomorphic

We conducted a cross-sectional study to determine the prevalence and species of gastro-intestinal helminths and haemoparasites in 100 chickens kept under extensive management systems in Ghana, West Africa. All the examined chickens (100%) were infected with gastro-intestinal helminths; a total of 18 species were detected. The species and their prevalences were: Acuaria hamulosa (25%), Allodapa suctoria (20%), Ascaridia galli (24%), Capillaria spp. (60%), Choanotaenia infundibulum (13%), Gongylonema ingluvicola (62%), Heterakis gallinarum (31%), H. isolonche (16%), Hymenolepis spp. (66%), Raillietina cesticillus (12%), R. echinobothrida (81%), R. tetragona (59%), Strongyloides avium (2%), Subulura strongylina (10%), Tetrameres fissispina (58%), Trichostronygylus tenuis (2%), and finally one unidentified acanthocephalan (1%) and one unidentified trematode (1%). Thirty-five per cent of the chickens were infected with the haemoparasites Aegyptinella pullorum and Plasmodium juxtanucleare (prevalences 9% and 27%, respectively). Association between chicken sex and prevalences was not significant. An over-dispersed distribution was seen for most of the helminth species.

Se realizó un estudio de los esquizontes exoeritrocíticos plasmáticos de Plasmodium juxtanucleare en gallinas sin raza definida, provenientes del município de Seropédica, estado de Rio de Janeiro, Brasil. Al examen sanguíneo se pudo observar en aves con alto índice de parasitémia eritrocítica (> 10%) formas esquizogónicas exoeritrocíticas libres, con un tamaño promedio de 7,492 ± 2,308 X 5,926 ± 2,048 µm; conteniendo una média de 15 ± 16,92 merozoítas. Estos esquizontes se presentaron con características pleomórficas de redondeado a oval, con merozoítos de núcleo basofílico. Se observó también, formas de esquizontes rotos con merozoítas libres. Estas y otras observaciones, reportadas y discutidas, hacen inferir que la cepa de P. juxtanucleare estudiada, realiza un ciclo paraeritrocítico. Se sugiere que P. juxtanucleare es una espécie de plasmodio, evolutivamente, intermediário entre los plasmodios de reptiles y los de mamíferos . A research about plasmatic exoerythrocytic meronts of the Plasmodium juxtanucleare in crossbred fowls from Seropédica municipality, Rio de Janeiro state, Brazil, was made. The hemoscopical exam of fowls with high erythrocytic parasitaemia (>10%) showed free exoerythrocytic meronts with an average size of 7.492 ± 2.308 X 5.926 ± 2.048 µm; containig about 15 ± 16.92 merozoites. The meronts showed to be pleomorphic characteristics from spherical to oval with merozoites of basophilic nucleus. They also showed torn meronts with free merozoites. All the observations reported and discussed about this subject make us conclude that P. juxtanucleare studied strain has a paraerythrocytic merogony. It suggests that P. juxtanucleare is an evolutionary plasmodia species intermediate between the plasmodia of reptiles and the plasmodia of mammals.

Foram feitas duas séries de experiências para verificar a possibilidade da infecção de mosquitos com o P. juxtanucleare. Na primeira foram utilizados 158 exemplares de Aedes aegypti e 152 de Culex quinquefasciatus, com resultados negativos. Na segunda série foram usados 15 espécimes de Aedes lepidus, com resultado negativo, e 80 de C. quinquefasciatus. Dêstes últimos apenas 3 se infectaram, não tendo sido possível transmitir a infecção a pintos inoculados. Êste culicíneo não é considerado transmissor habitual do plasmódio em estudo em vista do baixo índice de infectividade observado nas experiências. São brevemente discutidos os resultados do presente trabalho, inclusive o insucesso das inoculações em pintos.In order to determine the possibility of infecting mosquitoes with P. juxtanucleare two series of experiments were made. In the first series 158 Aëdes aegypti and 152 Culex quinquefasciatus were examined with negative results. In the second experiments 15 Aëdes lepidus and 80 C. quinquefasciatus were used. All Aëdes lepidus were negative. Theree Culex were found to have become experimentally infected. However, no infection was produced in chickens which were inoculated with salivary glands or stomach removed form the three mosquitoes. C. quinquefasciatus is not considered to be the common vector of P. juxtanucleare because of the very low infection index presented. The results, including those of the unsuccessful chicken inoculations, are briefly discussed.

1. The coccidial parasites (Eimeria acervulina and E. mitis) cause a recrudescence in chicks harbouring chronic infection with Plasmodium juxtanucleare.2. E. acervulina causes higher levels of parasitaemia in P. juxtanucleare infections in chicks if the coccidial infection is produced during the prepatent period of malaria, but it does not affect the length of patent and prepatent periods or raise the mortality rate among infected chicks.3. E. acervulina induces a severe fatal relapse in chicks harbouring latent P. gallinaceum infections.I wish to express my sincere thanks to Professor P.C.C. Garnham who supervised this work, and Dr R. Lainson who helped in identification, isolation of the coccidial parasites and the establishment of infection in clean laboratory-bred chicks.

INTRODUCTION THE species of Plasmodium known to occur naturally in chickens and cause malaria in that host are Plasmodium gallinaceum (Brumpt, 1935) and P. juxtanucleare (Versiani and Gomes, 1941). The former species has been recorded from fowls in Ceylon, Indo-China, Malaya and Sumatra. Natural outbreaks in fowls have been described (Crawford, 1945) in Ceylon. Occurrence of two naturally acquired infections in exotic fowls at the Izatnagar Poultry Farm, India, was described (Rao, Das and Ramnani, 1951Rao, Das and Ramnani, 1952). Plasmodium juxtanucleare has been described in chickens in Brazil by Versiani (1943) and in Mexico by Beltran (1943). The natural occurrence of a Plasmodium in chickens within the United States has apparently not been reported previously. OBSERVATIONS OF NATURAL INFECTIONS In June, 1959, during a survey of chicken parasites in Wisconsin, a Plasmodium-like organism was discovered in a blood smear from a 6-week-old chicken, one of a small flock maintained for research work …

Die Feinstruktur vonPlasmodium juxlanucleare wurde im Milzgewebe experimentell infizierter junger Huhnchen untersucht und die verschiedenen Veranderungen im Verlaufe des intrazellularen Entwicklungszyklus beschrieben.

SYNOPSIS. Blood films were examined from 1477 birds of Taiwan (193 species, 49 families). Haemoproteus Kruse was by far the commonest parasite, with Leucocytozoon Danilewski a not very close second. It is probable that some of the Haemoproteus infections represented new species, and 1 occurring in the Bamboo Partridge (Bambusicola thoracica sonorivox Gould) seemed characteristic enough to justify recognition as such; the name Haemoproteus bambusicolae sp. n. is proposed for this organism. Malaria was found in 77 birds, the greatest number of infections occurring in the Bamboo Partridge. Most of them were caused by Plasmodium juxtanucleare Versiani & Gomes, a pathogen of chickens, but a number were due to an undetermined species of Plasmodium. The Bamboo Partridge may be a reservoir host of the former. A few other identified species (P. rouxi Sergent & Sergent, P. hexamerium Huff, P. tenue Laveran & Mesnil) were seen, as well as some unidentified ones. Plasmodium tenue was seen in Garrulax canorus taewanus Swinhoe, a babbler: until now it was known only from the Pekin Robin (Leiothrix luteus Scopoli), also a babbler, in which we have found it extremely common. Sixty-four microfilarial infections were identified; they were especially frequent in the Button Quail (Turnix suscitator rostrata Swinhoe).

The numbers of malarial oocysts developing in individual, like mosquitoes fed concurrently on a single vertebrate malarial host were found to be distributed according to the negative binomial distribution in 169 experiments utilizing 6 species of Plasmodium, 6 species of mosquitoes and 3 species of vertebrate hosts. Dispersion constants ranged upward to 8.0, and mean clump sizes ranged upward to 298.4. The dispersion constant was demonstrated to be contingent on the species, strain and identity of the mosquito, the parasite and the vertebrate host; on the genetic state of the mosquito; and on the state of the infection in the vertebrate host. It was concluded that the concentration of oocyst production in particular mosquitoes was produced by varying levels and combinations of numerous factors associated with the parasite, the mosquito and the vertebrate host and that the pattern of oocyst distribution favors parasite survival and the maintenance of malaria in the field.

Plasmodium juxtanucleare was first described from domestic fowl in Brazil. Since that time similar parasites have been reported from Mexico, Uruguay, and Ceylon. Recently a morphologically similar form has been found widely distributed in the domestic fowl of Malaysia. The Malaysian strain of P. juxtanucleare is described and illustrated. The Asian strains are morphologically quite similar. However, some differences are noted between the Malaysian and the Brazilian strains. Versiani and Gomes (1941) described a malarial parasite from the domestic fowl (Gallus gallus) in Minas Gerais, Brazil which they named Plasmodium juxtanucleare. In the same year Beltran isolated a similar parasite from the same genus of fowl in Huixtla, Chiapas, Mexico. More detailed descriptions of this parasite along with observations on the frequently fatal course of experimental infections in chickens were published by the same authors 2 years later (Versiani and Gomes, 1943; Beltran, 1943). In 1945 Cassamagnaghi reported this parasite from domestic chickens in Uruguay. Ishiguro (1957) described a new species of malaria from Japanese domestic fowl which he named Plasmodium japonicum. This parasite had morphological characters similar to P. juxtanucleare but it was relatively nonpathogenic. Later, Dhanapala (1962) reported what he considered to be P. juxtanucleare from Ceylonese chickens. This latter parasite also produced a low mortality rate in chickens. Recently a parasite, indistinguishable from Dhanapala's Ceylon strain of P. juxtanucleare, has been found widely distributed as a generally nonlethal infection in the domestic fowl of the Malay states. This parasite has also been identified as P. juxtanucleare. The present paper describes and illustrates the Malaysian strain of this parasite. MATERIALS AND METHODS Source of the parasite During March-April of 1963, mosquitoes caught in monkey-baited net traps in the canopy of the Received for publication 1 March 1966. * Present address: Ontario Research Foundation, 43 Queen's Park, Toronto 5, Ontario, Canada. coastal mangrove swamp near Rantau Panjang, approximately 5 miles NW of Klang, State of Selangor, Malaysia, were examined for malarial parasites. In April, a Culex (Culex) sitiens Wiedmann, caught in one of the traps, was dissected and the salivary glands found to contain numerous sporozoites. Approximately one-half of the sporozoites were inoculated subcutaneously into a canary, the remainder intravenously into a chick. The former inoculation failed to produce an infection. The latter produced a patent parasitemia at 14 days. This parasite has subsequently been maintained in this laboratory by a succession of blood and sporozoite transfers. In June 1963 a Culex (Culex) annulus Theobald, taken in a hill forest region (Ulu Gombak) near Kuala Lumpur, was found to be infected with sporozoites morphologically similar to those mentioned above. When injected into a chick a patent infection resulted on day 12. The blood parasites were morphologically indistinguishable from those obtained at Rantau Panjang. On further study, various aspects of the biology of this parasite were found to be identical with those of the Rantau Panjang isolate and both isolations were considered to be the same parasite. Hatchery-reared chickens from the local market were used throughout the study. All birds were housed in mosquito-proof cages for a period of at least 2 weeks prior to their use. Their parasitefree status was confirmed by blood examinations prior to use. No natural infections with malaria were found in chickens from this source. Blood films were taken daily from either the brachial artery or vein. Routine preparations were stained with Giemsa stain at pH 7.2, and all descriptions, unless specifically noted, were made from these films. Pigment granules were more readily observed by staining alcohol-fixed films with thiazine red (5% thiazine red, 1% acetic acid in distilled water) for 3 min. Nuclear elements of both the host cell and the parasite remained unstained whereas the c toplasm of both stained light red. Pigment granules stood out clearly as blackish brown clumps. Measurements in the text are presented in the following way: average (range)/number of measurements. For example, the length of the schizonts is presented as 2.7 u (2 to 4)/39.

Some aspects of the life history of the Malaysian strain of Plasmodium juxtanucleare are presented. The periodicity of the parasite is believed to be an asynchronous, quotidian parasitemia with a long period of chronicity. Mature parasites are more common in visceral blood than in the peripheral circulation, indicating deep circulation schizogony. The parasite normally invades mature erythrocytes, with multiple infection of a cell uncommon except in instances of high parasitemia. The gametocytes are only slightly infective to mosquitoes during the first 30 days of patency, but subsequently become much more infective. Chronic infections remain highly infective to mosquitoes for as long as 198 days. The parasite could not be transmitted to canaries or young ducks. In the first paper of this series (Bennett and Warren, 1966) the asexual stages of the Malaysian strain of Plasmodium juxtanucleare were described, and in the second paper (Bennett et al., 1966) the development of the same parasite in the mosquito host was presented. The biology of P. juxtanucleare has been investigated to some extent by Al-Dabagh (1961) working with the Brazilian strain and by Dhanapala (1962) with the Ceylonese strain. This report is concerned with the biology of the Malaysian strain in the avian host with particular reference to periodicity of the asexual parasites in the peripheral blood, host cell preference, and infectivity of the gametocytes. In addition, this strain is compared with other known strains of the parasite and with other members of the subgenus Novyella. MATERIALS AND METHODS The parasite was maintained in the laboratory in young, hatchery-bred chickens by a combination of parasitized blood and sporozoite transfers. All birds were maintained in mosquito-proof cages. Subcutaneous, intravenous, and intraperitoneal routes were used for inoculations. Heparin was used as the anticoagulant for all parasitized blood transfers. Routine blood smears were made from the brachial vein at 0800 hr and stained with Giemsa stain at pH 7.2. Incisions through the body wall above the liver on anesthetized birds provided access to the liver for the hepatic smears. The incisions were closed with adhesive plaster and the birds treated with penicillin. The birds were then Received for publication 1 March 1966. * Present address: Ontario Research Foundation, 43 Queen's Park, Toronto 5, Ontario, Canada. anesthetized with ether, the incision opened, and smears made from the cut surface of the liver at desired intervals. Sporozoites for both routine and experimental transfers were obtained by dissecting the salivary glands from infected mosquitoes into Haye's saline. The glands were then crushed and the sporozoites collected in a mixture of 90% physiological saline10% serum (monkey or rabbit).