Chemical elements in sediments and in bivalve mollusks from estuarine regions in the south of Bahia State, northeast Brazil

Abstract Estuaries receive daily inputs of chemical elements which can impact the quality of water and sediment, as well as the health of biota. In addition to the sediment, bivalve mollusks have been used in the chemical monitoring of these systems. This study investigated the presence and contents of As, Ba, Cd, Co, Cr, Cu, Fe, Mn, Ni, Pb and Zn in superficial sediment and in bivalves (Crassostrea gasar, C. rhizophorae and Mytella guyanensis) from estuaries in the south / extreme south of Bahia State, northeast Brazil. The samples were evaluated with inductively coupled plasma optical emission spectrometry (ICP-OES, Varian 710). Except for Cd, all other elements were found in the samples, being that Co was exclusive in the sediment. The estuaries equivalent to sampling stations /1 - Valença, /2 - Taperoá, /3 - Ilhéus and /4 - Belmonte showed levels of metals compatibles with those established by the Brazilian legislation, however, the /5 - Santa Cruz Cabrália, in addition to the presence of As, presented a high level of Pb and Cu in C. gasar, which was attributed to the impacts of nautical activities in that locality.

Resumo Estuários recebem entradas diárias de elementos químicos, que podem impactar a qualidade de água e do sedimento, assim como a saúde da biota. Além do sedimento, moluscos bivalves têm sido utilizados no monitoramento químico desses sistemas. Neste estudo investigou-se a presença e os teores de As, Ba, Cd, Co, Cr, Cu, Fe, Mn, Ni, Pb e Zn no sedimento superficial e em bivalves (Crassostrea gasar, C. rhizophorae e Mytella guyanensis) de estuários do sul / extremo sul do estado da Bahia, Nordeste do Brasil. As amostras foram avaliadas por espectrometria de emissão óptica de plasma indutivamente acoplado (ICP-OES, Varian 710). Exceto Cd, todos os demais elementos foram encontrados nas amostras, sendo que Co foi exclusivo no sedimento. Os estuários equivalentes às estações amostrais /1 - Valença, /2 - Taperoá, /3 - Ilhéus e /4 - Belmonte mostraram níveis de metais compatíveis com os estabelecidos pela legislação brasileira, porém, a /5 - Santa Cruz Cabrália, além da presença de As, apresentou alto nível de Pb e de Cu em C. gasar, o que foi atribuído aos impactos por atividades náuticas nesse local.

Chemical elements in sediments and in bivalve mollusks from estuarine regions in the south of Bahia State, northeast Brazil / Elementos químicos em sedimentos e em moluscos bivalves de regiões estuarinas do sul do Estado da Bahia, Nordeste do Brasil

Estuaries receive daily inputs of chemical elements which can impact the quality of water and sediment, as well as the health of biota. In addition to the sediment, bivalve mollusks have been used in the chemical monitoring of these systems. This study investigated the presence and contents of As, Ba, Cd, Co, Cr, Cu, Fe, Mn, Ni, Pb and Zn in superficial sediment and in bivalves (Crassostrea gasar, C. rhizophorae and Mytella guyanensis) from estuaries in the south / extreme south of Bahia State, northeast Brazil. The samples were evaluated with inductively coupled plasma optical emission spectrometry (ICP-OES, Varian 710). Except for Cd, all other elements were found in the samples, being that Co was exclusive in the sediment. The estuaries equivalent to sampling stations #1 - Valença, #2 - Taperoá, #3 - Ilhéus and #4 - Belmonte showed levels of metals compatibles with those established by the Brazilian legislation, however, the #5 - Santa Cruz Cabrália, in addition to the presence of As, presented a high level of Pb and Cu in C. gasar, which was attributed to the impacts of nautical activities in that locality.

Estuários recebem entradas diárias de elementos químicos, que podem impactar a qualidade de água e do sedimento, assim como a saúde da biota. Além do sedimento, moluscos bivalves têm sido utilizados no monitoramento químico desses sistemas. Neste estudo investigou-se a presença e os teores de As, Ba, Cd, Co, Cr, Cu, Fe, Mn, Ni, Pb e Zn no sedimento superficial e em bivalves (Crassostrea gasar, C. rhizophorae e Mytella guyanensis) de estuários do sul / extremo sul do estado da Bahia, Nordeste do Brasil. As amostras foram avaliadas por espectrometria de emissão óptica de plasma indutivamente acoplado (ICP-OES, Varian 710). Exceto Cd, todos os demais elementos foram encontrados nas amostras, sendo que Co foi exclusivo no sedimento. Os estuários equivalentes às estações amostrais #1 - Valença, #2 - Taperoá, #3 - Ilhéus e #4 - Belmonte mostraram níveis de metais compatíveis com os estabelecidos pela legislação brasileira, porém, a #5 - Santa Cruz Cabrália, além da presença de As, apresentou alto nível de Pb e de Cu em C. gasar, o que foi atribuído aos impactos por atividades náuticas nesse local.

Chemical elements in sediments and in bivalve mollusks from estuarine regions in the south of Bahia State, northeast Brazil / Elementos químicos em sedimentos e em moluscos bivalves de regiões estuarinas do sul do Estado da Bahia, Nordeste do Brasil

Estuaries receive daily inputs of chemical elements which can impact the quality of water and sediment, as well as the health of biota. In addition to the sediment, bivalve mollusks have been used in the chemical monitoring of these systems. This study investigated the presence and contents of As, Ba, Cd, Co, Cr, Cu, Fe, Mn, Ni, Pb and Zn in superficial sediment and in bivalves (Crassostrea gasar, C. rhizophorae and Mytella guyanensis) from estuaries in the south / extreme south of Bahia State, northeast Brazil. The samples were evaluated with inductively coupled plasma optical emission spectrometry (ICP-OES, Varian 710). Except for Cd, all other elements were found in the samples, being that Co was exclusive in the sediment. The estuaries equivalent to sampling stations #1 - Valença, #2 - Taperoá, #3 - Ilhéus and #4 - Belmonte showed levels of metals compatibles with those established by the Brazilian legislation, however, the #5 - Santa Cruz Cabrália, in addition to the presence of As, presented a high level of Pb and Cu in C. gasar, which was attributed to the impacts of nautical activities in that locality.(AU)

Estuários recebem entradas diárias de elementos químicos, que podem impactar a qualidade de água e do sedimento, assim como a saúde da biota. Além do sedimento, moluscos bivalves têm sido utilizados no monitoramento químico desses sistemas. Neste estudo investigou-se a presença e os teores de As, Ba, Cd, Co, Cr, Cu, Fe, Mn, Ni, Pb e Zn no sedimento superficial e em bivalves (Crassostrea gasar, C. rhizophorae e Mytella guyanensis) de estuários do sul / extremo sul do estado da Bahia, Nordeste do Brasil. As amostras foram avaliadas por espectrometria de emissão óptica de plasma indutivamente acoplado (ICP-OES, Varian 710). Exceto Cd, todos os demais elementos foram encontrados nas amostras, sendo que Co foi exclusivo no sedimento. Os estuários equivalentes às estações amostrais #1 - Valença, #2 - Taperoá, #3 - Ilhéus e #4 - Belmonte mostraram níveis de metais compatíveis com os estabelecidos pela legislação brasileira, porém, a #5 - Santa Cruz Cabrália, além da presença de As, apresentou alto nível de Pb e de Cu em C. gasar, o que foi atribuído aos impactos por atividades náuticas nesse local.(AU)

In vitro culture of parasitic stages of Haemonchus contortus / Cultivo in vitro de estádios parasitários de Haemonchus contortus

Haemonchus contortus is a constraint to sheep production. Seeking to reduce the use of hosts and produce parasitic stages in large-scale, a 42-day in vitro culture protocol of H. contortus third-stage larvae was optimized using Dulbecco's modified Eagle's medium (DMEM). In cell-free culture, larvae were maintained at 39.6°C, in acidic media (pH 6.1) for 3 or 6 days with Δ4-dafachronic acid followed by DMEM pH 7.4 supplemented or not with Fildes' reagent. In DMEM pH 7.4 at 37°C, supplementation with Caco-2 cells was compared to Fildes. On Day 14, fourth-stage larvae (L4) development rates in acidic media supplemented (86.8-88.4%) or not (74.4-77.8%) with Fildes and in Caco-2 cell co-culture (92.6%) were similar, and superior to DMEM pH 7.4 with Fildes (0.0%). On Day 21, Caco-2 cell co-culture resulted in higher larvae differentiation (25.0%) and lower degeneration (13.9%) compared to acidic media (1.5-8.1% and 48.6-69.9%, respectively). This is the first report of prolonged in vitro culture of H. contortus larvae using commercial media in co-culture with Caco-2 cells. Although no progression to the adult stage, Caco-2 cell co-culture resulted in morphological differentiation of H. contortus L4 and larval viability for up to 28 days.(AU)

Haemonchus contortus provoca grandes prejuízos à ovinocultura. Visando reduzir a utilização de ovinos hospedeiros e produzir estágios parasitários em larga escala, um protocolo para o cultivo in vitro por 42 dias de larvas de terceiro estádio de H. contortus foi realizado em meio Eagle, modificado por Dulbecco (DMEM). No cultivo sem células, as larvas foram mantidas a 39,6°C e incubadas em DMEM ácido (pH 6,1) por 3 ou 6 dias com 4Δ-ácido dafacrônico seguido por DMEM pH 7,4 suplementado ou não com reagente de Fildes. Em DMEM pH 7,4 a 37°C, a suplementação com células Caco-2 foi comparada à suplementação com Fildes. No Dia 14, as taxas de desenvolvimento até o quarto estádio larvar (L4) foram similares em meio ácido sem células suplementado (86,8-88,4%) ou não (74,4-77,8%) com Fildes e em co-cultura com células Caco-2 (92,6%), e superiores ao desenvolvimento em DMEM pH 7,4 com Fildes (0,0%). No Dia 21, o co-cultivo com células Caco-2 resultou em maior diferenciação (25,0%) e menor degeneração (13,9%) das larvas em comparação ao meio ácido (1,5-8,1% e 48,6-69,9%, respectivamente). Este é o primeiro relato de cultivo in vitro prolongado de H. contortus em meio comercial em co-cultivo com células Caco-2. Apesar da ausência de progressão até o estágio adulto, o co-cultivo com células Caco-2 resultou em diferenciação de L4 e manutenção da viabilidade das larvas de H. contortus por até 28 dias in vitro.(AU)

Chemical elements in sediments and in bivalve mollusks from estuarine regions in the south of Bahia State, northeast Brazil / Elementos químicos em sedimentos e em moluscos bivalves de regiões estuarinas do sul do Estado da Bahia, Nordeste do Brasil

Abstract Estuaries receive daily inputs of chemical elements which can impact the quality of water and sediment, as well as the health of biota. In addition to the sediment, bivalve mollusks have been used in the chemical monitoring of these systems. This study investigated the presence and contents of As, Ba, Cd, Co, Cr, Cu, Fe, Mn, Ni, Pb and Zn in superficial sediment and in bivalves (Crassostrea gasar, C. rhizophorae and Mytella guyanensis) from estuaries in the south / extreme south of Bahia State, northeast Brazil. The samples were evaluated with inductively coupled plasma optical emission spectrometry (ICP-OES, Varian 710). Except for Cd, all other elements were found in the samples, being that Co was exclusive in the sediment. The estuaries equivalent to sampling stations #1 - Valença, #2 - Taperoá, #3 - Ilhéus and #4 - Belmonte showed levels of metals compatibles with those established by the Brazilian legislation, however, the #5 - Santa Cruz Cabrália, in addition to the presence of As, presented a high level of Pb and Cu in C. gasar, which was attributed to the impacts of nautical activities in that locality.

Resumo Estuários recebem entradas diárias de elementos químicos, que podem impactar a qualidade de água e do sedimento, assim como a saúde da biota. Além do sedimento, moluscos bivalves têm sido utilizados no monitoramento químico desses sistemas. Neste estudo investigou-se a presença e os teores de As, Ba, Cd, Co, Cr, Cu, Fe, Mn, Ni, Pb e Zn no sedimento superficial e em bivalves (Crassostrea gasar, C. rhizophorae e Mytella guyanensis) de estuários do sul / extremo sul do estado da Bahia, Nordeste do Brasil. As amostras foram avaliadas por espectrometria de emissão óptica de plasma indutivamente acoplado (ICP-OES, Varian 710). Exceto Cd, todos os demais elementos foram encontrados nas amostras, sendo que Co foi exclusivo no sedimento. Os estuários equivalentes às estações amostrais #1 - Valença, #2 - Taperoá, #3 - Ilhéus e #4 - Belmonte mostraram níveis de metais compatíveis com os estabelecidos pela legislação brasileira, porém, a #5 - Santa Cruz Cabrália, além da presença de As, apresentou alto nível de Pb e de Cu em C. gasar, o que foi atribuído aos impactos por atividades náuticas nesse local.

Calves with arthritis - changes in antioxidant parameters

Background: In a healthy organism, oxidants and antioxidants are in balance. However, in cases such as inflammation, infection, and stress, this balance is disrupted in favor of oxidants, creating oxidative stress that can cause damage to cells or tissues. It is known that oxidative stress plays a role in the pathogenesis of many diseases. Determination of oxidant and antioxidant balance, especially in inflammatory diseases, plays an important role in elucidating the pathogenesis of the disease and developing treatment strategies. This study, it was aimed to reveal the oxidant status in inflammatory disease of calves with septic and aseptic arthritis. Materials, Methods & Results: The material of the study consisted of 21 calves up to 2 months old, of different races and genders, 14 (9 male, 5 female) with arthritis and 7 healthy (control, 5 male, 2 female). Of the calves with arthritis, 11 were septic and 3 were acute aseptic. In the calves with arthritis, the affected joint or joints were determined by clinical examinations. By palpating the joints, swelling, local temperature increase, tension in the joint capsule, presence of pain, and the presence and severity of lameness were examined. The color, clarity, viscosity, odor, and clot formation of the synovial fluid were examined and determined to be septic or aseptic. To determine the antioxidant status, the levels of malondialdehyde (MDA), which is the most important oxidative stress marker, and superoxide dismutase (SOD), gluta-thione peroxidase (GSH-Px), and catalase (CAT), which are the enzymatic antioxidant enzymes, were measured spectro-photometrically in serum samples. Vitamin E, C, and A levels, which are nonenzymatic antioxidants, were also measured colorimetrically. In the clinical examination, lameness was detected in the relevant extremity of all patients with arthritis. In the macroscopic examination of the synovial fluids taken from animals with arthritis, the colors of the synovial fluids varied between yellow and yellow tones in 11 cases; in 3 cases, it was determined that they were red and brown. It was observed that the colors of the synovial fluids were transparent in the subjects in the control group. It was observed that the synovial fluid clarity of the calves with arthritis was lost, with severe turbidity (+++) in 3 cases, moderately turbid (++) in 6 cases, slightly turbid (+) in 2 cases, and clear (-) in 3 cases. It was observed that the viscosity of synovial fluid taken from calves with arthritis decreased in varying degrees according to the severity of the disease, severe (+++) in 5 cases, moderately decreased (++) in 4 cases, slightly decreased (+) in 2 cases, and normal in 3 cases. It was determined that the viscosity of the synovial fluid taken from the calves in the control group was normal. There was a statistically significant difference between the groups in terms of MDA (P < 0.01), SOD (P < 0.01), GSH-Px (P < 0.05), vitamin E (P < 0.001), and vitamin C (P < 0.01), while MDA levels increased in calves with arthritis, SOD and GSH-Px activities and vitamin E and C levels decreased significantly. Although there was no statistically significant difference in CAT (P > 0.05) enzyme activity, it was determined that it was at a lower level in calves with arthritis, and there was no significant difference be-tween the groups in terms of vitamin A (P > 0.05). Discussion: According to the results of the study, there is an increase in oxidative stress and a decrease in antioxidant status in calves with arthritis. It is thought that these changes may be due to efforts to reduce tissue damage by reducing lipid peroxidation. As a result, it was determined that oxidant and antioxidant balance was impaired in calves with arthritis, and oxidative stress and lipid peroxidation developed due to the increase in free radicals. It is thought that giving additional antioxidants to the calves may contribute to the recovery of the disease and reduce treatment costs.

Gastrointestinal parasites in captive and free-living wild birds in Goiania Zoo

Abstract Helminths are endoparasites that infect a variety of bird species. Endoparasite infections can cause severe diseases, including kill captive avian hosts and represents a problem to maintenance and conservation. The eggs and larval forms of these parasites are usually eliminated in the host feces. The main interest of this study is to report the occurrence of eggs and oocysts in feces from captive wild birds in Goiânia Zoo and free-living birds in its surroundings. The fecal samples were subjected to parasitological examination to identify the presence of helminths and to classify their eggs based on morphological characteristics. Eggs of nematode parasites (Positive/N) were identified as Ascaridia spp. in Brotogeris chiriri (2/2), Dromaius novaehollandiae (1/1) and Rhea americana (2/2); Ascarididae in Pavo cristatus nigripensis (1/1); Capillaria spp. in Ara chloropterus (1/1) and Penelope jacucaca (1/1); Capillaria plagiaticia in Anodorhynchus hyacinthinus (1/2) and Ara spp. (2/2); Capillaria venusta in Rhamphastus tucanus (1/2); and unidentified nematode eggs from Amazona amazonica (2/2). Eimeria spp. oocyst was founded in feces from A. hyacinthinus (1/2). Considering free-living birds, Diphyllobothrium spp. eggs were founded in Ardea alba (2/2) and Nycticorax nycticorax (2/3). Co-infection of Eustrongylides spp. was founded in A. alba (1/2). This is the first occurrence of: Ascaridia spp. parasitizing B. chiriri; and C. venusta parasitizing R. tucanus. In conclusion, the helminth eggs found in the abovementioned host bird species are consistent with those reported in the literature, and treatment and control protocols were based on their identification.

Resumo Helmintos são endoparasitas que infectam uma variedade de espécies de aves. As infecções por endoparasitas podem causar doenças graves, podendo matar aves em cativeiro e representam um problema de manutenção e conservação. Os ovos e as formas larvais desses parasitas são geralmente eliminados nas fezes do hospedeiro. O principal objetivo deste estudo é relatar a ocorrência de ovos e oocistos em fezes de aves silvestres em cativeiro no Zoológico de Goiânia e em aves de vida livre em seu entorno. As amostras fecais foram submetidas a exame parasitológico para identificação da presença de helmintos e classificação dos ovos com base nas características morfológicas. Ovos de nematóides (Positivo/N) foram identificados como Ascaridia spp. em Brotogeris chiriri (2/2), Dromaius novaehollandiae (1/1) e Rhea americana (2/2); Ascarididae em Pavo cristatus nigripensis (1/1); Capillaria spp. em Ara chloropterus (1/1) e Penelope jacucaca (1/1); Capillaria plagiaticia em Anodorhynchus hyacinthinus (1/2) e Ara spp. (2/2); Capillaria venusta em Rhamphastus tucanus (1/2); e ovos de nematóide não identificado de Amazona amazonica (2/2). Oocistos de Eimeria spp. foram encontrados nas fezes de A. hyacinthinus (1/2). Considerando pássaros de vida livre, ovos de Diphyllobothrium spp. foram encontrados em Ardea alba (2/2) e Nycticorax nycticorax (2/3). Coinfecção de Eustrongylides spp. foi encontradada em A. alba (1/2). Esta é a primeira ocorrência de: Ascaridia spp. parasitando B. chiriri; e C. venusta parasitando R. tucanus. Em conclusão, os ovos de helmintos encontrados nas espécies de aves hospedeiras acima mencionadas são consistentes com os relatados na literatura, e os protocolos de tratamento e controle foram baseados em sua identificação.

Gastrointestinal parasites in captive and free-living wild birds in Goiania Zoo / Parasitas gastrointestinais em aves silvestres em cativeiro e de vida livre no Zoológico de Goiânia

Helminths are endoparasites that infect a variety of bird species. Endoparasite infections can cause severe diseases, including kill captive avian hosts and represents a problem to maintenance and conservation. The eggs and larval forms of these parasites are usually eliminated in the host feces. The main interest of this study is to report the occurrence of eggs and oocysts in feces from captive wild birds in Goiânia Zoo and free-living birds in its surroundings. The fecal samples were subjected to parasitological examination to identify the presence of helminths and to classify their eggs based on morphological characteristics. Eggs of nematode parasites (Positive/N) were identified as Ascaridia spp. in Brotogeris chiriri (2/2), Dromaius novaehollandiae (1/1) and Rhea americana (2/2); Ascarididae in Pavo cristatus nigripensis (1/1); Capillaria spp. in Ara chloropterus (1/1) and Penelope jacucaca (1/1); Capillaria plagiaticia in Anodorhynchus hyacinthinus (1/2) and Ara spp. (2/2); Capillaria venusta in Rhamphastus tucanus (1/2); and unidentified nematode eggs from Amazona amazonica (2/2). Eimeria spp. oocyst was founded in feces from A. hyacinthinus (1/2). Considering free-living birds, Diphyllobothrium spp. eggs were founded in Ardea alba (2/2) and Nycticorax nycticorax (2/3). Co-infection of Eustrongylides spp. was founded in A. alba (1/2). This is the first occurrence of: Ascaridia spp. parasitizing B. chiriri; and C. venusta parasitizing R. tucanus. In conclusion, the helminth eggs found in the abovementioned host bird species are consistent with those reported in the literature, and treatment and control protocols were based on their identification.

Helmintos são endoparasitas que infectam uma variedade de espécies de aves. As infecções por endoparasitas podem causar doenças graves, podendo matar aves em cativeiro e representam um problema de manutenção e conservação. Os ovos e as formas larvais desses parasitas são geralmente eliminados nas fezes do hospedeiro. O principal objetivo deste estudo é relatar a ocorrência de ovos e oocistos em fezes de aves silvestres em cativeiro no Zoológico de Goiânia e em aves de vida livre em seu entorno. As amostras fecais foram submetidas a exame parasitológico para identificação da presença de helmintos e classificação dos ovos com base nas características morfológicas. Ovos de nematóides (Positivo/N) foram identificados como Ascaridia spp. em Brotogeris chiriri (2/2), Dromaius novaehollandiae (1/1) e Rhea americana (2/2); Ascarididae em Pavo cristatus nigripensis (1/1); Capillaria spp. em Ara chloropterus (1/1) e Penelope jacucaca (1/1); Capillaria plagiaticia em Anodorhynchus hyacinthinus (1/2) e Ara spp. (2/2); Capillaria venusta em Rhamphastus tucanus (1/2); e ovos de nematóide não identificado de Amazona amazonica (2/2). Oocistos de Eimeria spp. foram encontrados nas fezes de A. hyacinthinus (1/2). Considerando pássaros de vida livre, ovos de Diphyllobothrium spp. foram encontrados em Ardea alba (2/2) e Nycticorax nycticorax (2/3). Coinfecção de Eustrongylides spp. foi encontradada em A. alba (1/2). Esta é a primeira ocorrência de: Ascaridia spp. parasitando B. chiriri; e C. venusta parasitando R. tucanus. Em conclusão, os ovos de helmintos encontrados nas espécies de aves hospedeiras acima mencionadas são consistentes com os relatados na literatura, e os protocolos de tratamento e controle foram baseados em sua identificação.

Gastrointestinal parasites in captive and free-living wild birds in Goiania Zoo / Parasitas gastrointestinais em aves silvestres em cativeiro e de vida livre no Zoológico de Goiânia

Helminths are endoparasites that infect a variety of bird species. Endoparasite infections can cause severe diseases, including kill captive avian hosts and represents a problem to maintenance and conservation. The eggs and larval forms of these parasites are usually eliminated in the host feces. The main interest of this study is to report the occurrence of eggs and oocysts in feces from captive wild birds in Goiânia Zoo and free-living birds in its surroundings. The fecal samples were subjected to parasitological examination to identify the presence of helminths and to classify their eggs based on morphological characteristics. Eggs of nematode parasites (Positive/N) were identified as Ascaridia spp. in Brotogeris chiriri (2/2), Dromaius novaehollandiae (1/1) and Rhea americana (2/2); Ascarididae in Pavo cristatus nigripensis (1/1); Capillaria spp. in Ara chloropterus (1/1) and Penelope jacucaca (1/1); Capillaria plagiaticia in Anodorhynchus hyacinthinus (1/2) and Ara spp. (2/2); Capillaria venusta in Rhamphastus tucanus (1/2); and unidentified nematode eggs from Amazona amazonica (2/2). Eimeria spp. oocyst was founded in feces from A. hyacinthinus (1/2). Considering free-living birds, Diphyllobothrium spp. eggs were founded in Ardea alba (2/2) and Nycticorax nycticorax (2/3). Co-infection of Eustrongylides spp. was founded in A. alba (1/2). This is the first occurrence of: Ascaridia spp. parasitizing B. chiriri; and C. venusta parasitizing R. tucanus. In conclusion, the helminth eggs found in the abovementioned host bird species are consistent with those reported in the literature, and treatment and control protocols were based on their identification.

Helmintos são endoparasitas que infectam uma variedade de espécies de aves. As infecções por endoparasitas podem causar doenças graves, podendo matar aves em cativeiro e representam um problema de manutenção e conservação. Os ovos e as formas larvais desses parasitas são geralmente eliminados nas fezes do hospedeiro. O principal objetivo deste estudo é relatar a ocorrência de ovos e oocistos em fezes de aves silvestres em cativeiro no Zoológico de Goiânia e em aves de vida livre em seu entorno. As amostras fecais foram submetidas a exame parasitológico para identificação da presença de helmintos e classificação dos ovos com base nas características morfológicas. Ovos de nematóides (Positivo/N) foram identificados como Ascaridia spp. em Brotogeris chiriri (2/2), Dromaius novaehollandiae (1/1) e Rhea americana (2/2); Ascarididae em Pavo cristatus nigripensis (1/1); Capillaria spp. em Ara chloropterus (1/1) e Penelope jacucaca (1/1); Capillaria plagiaticia em Anodorhynchus hyacinthinus (1/2) e Ara spp. (2/2); Capillaria venusta em Rhamphastus tucanus (1/2); e ovos de nematóide não identificado de Amazona amazonica (2/2). Oocistos de Eimeria spp. foram encontrados nas fezes de A. hyacinthinus (1/2). Considerando pássaros de vida livre, ovos de Diphyllobothrium spp. foram encontrados em Ardea alba (2/2) e Nycticorax nycticorax (2/3). Coinfecção de Eustrongylides spp. foi encontradada em A. alba (1/2). Esta é a primeira ocorrência de: Ascaridia spp. parasitando B. chiriri; e C. venusta parasitando R. tucanus. Em conclusão, os ovos de helmintos encontrados nas espécies de aves hospedeiras acima mencionadas são consistentes com os relatados na literatura, e os protocolos de tratamento e controle foram baseados em sua identificação.

Gastrointestinal parasites in captive and free-living wild birds in Goiania Zoo / Parasitas gastrointestinais em aves silvestres em cativeiro e de vida livre no Zoológico de Goiânia

Helminths are endoparasites that infect a variety of bird species. Endoparasite infections can cause severe diseases, including kill captive avian hosts and represents a problem to maintenance and conservation. The eggs and larval forms of these parasites are usually eliminated in the host feces. The main interest of this study is to report the occurrence of eggs and oocysts in feces from captive wild birds in Goiânia Zoo and free-living birds in its surroundings. The fecal samples were subjected to parasitological examination to identify the presence of helminths and to classify their eggs based on morphological characteristics. Eggs of nematode parasites (Positive/N) were identified as Ascaridia spp. in Brotogeris chiriri (2/2), Dromaius novaehollandiae (1/1) and Rhea americana (2/2); Ascarididae in Pavo cristatus nigripensis (1/1); Capillaria spp. in Ara chloropterus (1/1) and Penelope jacucaca (1/1); Capillaria plagiaticia in Anodorhynchus hyacinthinus (1/2) and Ara spp. (2/2); Capillaria venusta in Rhamphastus tucanus (1/2); and unidentified nematode eggs from Amazona amazonica (2/2). Eimeria spp. oocyst was founded in feces from A. hyacinthinus (1/2). Considering free-living birds, Diphyllobothrium spp. eggs were founded in Ardea alba (2/2) and Nycticorax nycticorax (2/3). Co-infection of Eustrongylides spp. was founded in A. alba (1/2). This is the first occurrence of: Ascaridia spp. parasitizing B. chiriri; and C. venusta parasitizing R. tucanus. In conclusion, the helminth eggs found in the abovementioned host bird species are consistent with those reported in the literature, and treatment and control protocols were based on their identification.(AU)

Helmintos são endoparasitas que infectam uma variedade de espécies de aves. As infecções por endoparasitas podem causar doenças graves, podendo matar aves em cativeiro e representam um problema de manutenção e conservação. Os ovos e as formas larvais desses parasitas são geralmente eliminados nas fezes do hospedeiro. O principal objetivo deste estudo é relatar a ocorrência de ovos e oocistos em fezes de aves silvestres em cativeiro no Zoológico de Goiânia e em aves de vida livre em seu entorno. As amostras fecais foram submetidas a exame parasitológico para identificação da presença de helmintos e classificação dos ovos com base nas características morfológicas. Ovos de nematóides (Positivo/N) foram identificados como Ascaridia spp. em Brotogeris chiriri (2/2), Dromaius novaehollandiae (1/1) e Rhea americana (2/2); Ascarididae em Pavo cristatus nigripensis (1/1); Capillaria spp. em Ara chloropterus (1/1) e Penelope jacucaca (1/1); Capillaria plagiaticia em Anodorhynchus hyacinthinus (1/2) e Ara spp. (2/2); Capillaria venusta em Rhamphastus tucanus (1/2); e ovos de nematóide não identificado de Amazona amazonica (2/2). Oocistos de Eimeria spp. foram encontrados nas fezes de A. hyacinthinus (1/2). Considerando pássaros de vida livre, ovos de Diphyllobothrium spp. foram encontrados em Ardea alba (2/2) e Nycticorax nycticorax (2/3). Coinfecção de Eustrongylides spp. foi encontradada em A. alba (1/2). Esta é a primeira ocorrência de: Ascaridia spp. parasitando B. chiriri; e C. venusta parasitando R. tucanus. Em conclusão, os ovos de helmintos encontrados nas espécies de aves hospedeiras acima mencionadas são consistentes com os relatados na literatura, e os protocolos de tratamento e controle foram baseados em sua identificação.(AU)

Changes in somatic cell count, composition and cytokine levels in milk from cows with mastitis due to mixed infections

Mastitis is a mammary gland inflammation that is very common worldwide, mostly caused by bacteria, and causes enormous economic losses. Many microorganisms cause this disease. The most common causes of mastitis by these microorganisms are Staphylococcus aureus (S. aureus), Escherichia coli (E. coli) and Streptococcus agalactiae (S. agalactiae). The anti-inflammatory properties of transforming growth factor (TGF)-ß include: 1) limiting interferon (IFN)-γ production; 2) increasing the expression of the interleukine (IL)-1 receptor antagonist; 3) inhibiting macrophage production of chemokines, pro-inflammatory cytokines, nitric oxide, and reactive oxygen intermediates; and 4) increasing macrophage clearance of bacterial debris and damaged parenchymal cells. It is stated that cytokines and milk composition change in case of mastitis. In this study, it was aimed to reveal the changes in milk TGF-ß1 and Tumor necrosis factor (TNF)-α concentrations and milk composition in mixed infections caused by three pathogens causing mastitis. In this study, milk samples from 90 cows were divided into 5 groups. Tumor necrosis factor (TNF)-α and TGF-ß1 concentrations and milk composition were determined in these milk samples. The California Mastitis Test (CMT) was applied to the cows included in the study and scoring was done. According to the CMT results of the milk samples taken, CMT(-) cows were included in group 1 (n = 22). Those with the CMT(+) were sent to the microbiology laboratory for analysis within 2 h. After the bacteria was determined, combination groupings were formed. Group 2 (n = 17), in which S. aureus and E. coli grew together, group 3 (n = 21), in which S. aureus and S. agalactiae grew together, group 4 (n = 8), in which S. agalactiae and E. coli grew together in milk samples, and milk samples without any bacterial growth in CMT (+) formed group 5 (n = 22), respectively. Somatic cell count was measured with the DeLaval Cell Counter® (Cell Counter DCC) device. Mineral matter, fat, protein, lactose, electrical conductivity and specific gravity were measured in milk samples using Lactoscan Milk Analyzer (Milkotronic/EUROPE). Milk samples were then stored at -80°C to measure TGF-ß1 and TNF-α. Tumor necrosis factor-α and TGF-ß1 concentrations in milk samples were measured using ELISA kits (Sunred Biological Technology). Changes in milk TNF-α and TGF-ß1 concentration and milk composition were determined in milk samples with mastitis caused by mixed infection. The TNF-α concentration of group 4 was higher than the other groups. On the other hand, the highest concentration of TGF-ß1 was found in group 2. While the number of somatic cells in group 1 was lower than in groups 2, 3, and 4, there was no statistical difference between groups 1 and 5. The lowest milk fat ratio was found in group 1, and it was found to be statistically lower than groups 2, 3, and 4. While the rate of solid-non-fat of group 1 increased compared to groups 2 and 3, the highest protein ratio was found in groups 1 and 5. There was no difference between the 5 groups in terms of mineral matter ratios. While the specific gravity was highest in group 1, there was no statistical difference between the other 4 groups. Overall, it was concluded that there was an increase in TNF-α and TGF-ß1 concentrations and a change in milk composition in samples with bacterial growth.(AU)

Proposal of an iELISA for Mycoplasma bovis diagnosis in dairy cattle and associated risk factors / [Proposta de um iELISA para diagnóstico de Mycoplasma bovis em bovinos leiteiros e fatores de risco associados]

Mycoplasma bovis is a highly contagious agent associated with several pathologies in cattle. The detection of reactive antibodies to M. bovis by Indirect Enzyme-Linked Immunosorbent Assay (iELISA) identifies if there was an exposure to the microorganism. The current study aimed to optimize an iELISA from M. bovis total cell antigen, applying it to bovine serum samples, and to evaluate risk factors. Serum samples were obtained from 400 cows from 17 herds from Southeast Brazil. In the optimization of iELISA, the following was established: 2 µg/mL of antigen, sera dilution 1:300, and conjugate dilution 1:15000. The frequency was 62.3% (249/400) of reactive animals and 100% (17/17) of reactive herds. Risk factors were: herds with more than 100 animals (OR= 3.1; CI= 95%); Holstein breed (OR= 72.5; CI= 95%); cows (OR= 29.7; CI= 95%); intensive breeding system (OR= 3.3; CI= 95%); associated small ruminant production (OR= 4.4; CI= 95%); milk production above 500L (OR= 2.9; CI= 95%); no quarantine (OR= 1.5; CI= 95%); mechanical milking (OR= 5.5; CI= 95%) and cases of mastitis (OR= 5.5; CI= 95%). The proposed iELISA was able to detect antibodies reactive to M. bovis in bovine serum. Knowledge of these risk factors can assist in the implementation of prophylactic measures.(AU)

Mycoplasma bovis é um agente altamente contagioso relacionado a várias patologias em bovinos. A detecção de anticorpos reativos a M. bovis por Ensaio de Imunoadsorção Enzimática Indireto (iELISA) identifica se houve exposição ao microrganismo. O presente estudo teve como objetivo otimizar um iELISA de antígeno celular total de M. bovis, aplicando-o a amostras de soro bovino, bem como avaliar fatores de risco. Amostras de soro foram obtidas de 400 vacas de 17 rebanhos da Região Sudeste do Brasil. Na otimização do iELISA foram obtidos: 2µg/mL de antígeno, diluição dos soros 1:300 e do conjugado 1:15000. A frequência de animais reativos foi de 62,3% (249/400) e de 100% (17/17) para os rebanhos. Os fatores de risco foram: rebanhos com mais de 100 animais (OR= 3,1; IC= 95%); raça Holandesa (OR= 72,5;IC= 95%); vacas (OR= 29,7;IC= 95%); sistema intensivo (OR= 3,3; C= 95%); produção de pequenos ruminantes (OR= 4,4;IC=95%); produção de leite acima de 500L (OR= 2,9;IC= 95%); sem quarentena (OR= 1,5;IC= 95%); ordenha mecânica (OR= 5,5;IC= 95%) e casos de mastite (OR= 5,5;IC= 95%). O iELISA proposto foi capaz de detectar anticorpos reativos a M. bovis no soro bovino. O conhecimento desses fatores de risco pode auxiliar na implementação de medidas profiláticas.(AU)

Resíduo do processamento do milho doce em diferentes níveis e frequências para bovinos confinados / Sweet corn residue processed offered at different levels and frequencies for feedlot cattle

Objetivou-se avaliar o desempenho de bovinos de corte confinados recebendo diferentes níveis e frequências do resíduo do processamento do milho doce como volumoso. Os animais foram divididos em cinco grupos, totalizando 30 animais por tratamento: 1) relação volumoso: concentrado (V:C) de 50:50 do início ao final do experimento; 2) 30 dias com 100% V, seguidos de 30 dias com relação V:C de 50:50, e posteriormente relação V:C de 30:70 até o final do período experimental; 3) 45 dias iniciais do confinamento com 100% de volumoso, e posteriormente relação V:C de 50:50 até o final do período experimental; 4) 30 dias iniciais do confinamento com 100% de volumoso, e posteriormente relação V:C de 50:50 até o final do período experimental; 5) 100% de volumoso. O fornecimento exclusivo de volumoso durante o período total do confinamento proporcionou aos animais os menores peso vivo final, ganho médio diário e consumo de matéria seca, porém, apresentaram a melhor conversão alimentar entre os tratamentos avaliados e menor custo por arroba produzida. A inclusão de concentrado proporcionou, de maneira geral, maior desempenho dos animais, porém, aumento dos custos de produção.

Feedlot cattle performance was evaluated upon diets offering different levels and frequencies of sweet corn residue as source of roughage. The animals were divided into five groups, in a total of 30 animals per treatment. 1) roughage:grain ratio (R:G) of 50:50 from the start to the end of experiment , 2) 30 days with 100% roughage, followed by 30 days with R:G ratio of 50:50, and then R:G ratio of 30:70 until the end of the experimental period; 3) first 45 days at feedlot condition with 100% of roughage and then R:G ratio of 50:50 until the end of experimental period; 4) 30 days at start of feedlot with 100% of roughage, and then a ratio of 50:50 until the end of experimental period; 5) 100% of roughage. The lowest body weight, average daily gain and dry matter intake were achieved by animals receiving only roughage during the total period of feedlot; however, these animals presented the best feed conversion between evaluated treatments and lowest cost per pound produced. The concentrate inclusion offered, in general, highest animal performance, but with high-cost production.

Resíduo do processamento do milho doce em diferentes níveis e frequências para bovinos confinados / Sweet corn residue processed offered at different levels and frequencies for feedlot cattle

Objetivou-se avaliar o desempenho de bovinos de corte confinados recebendo diferentes níveis e frequências do resíduo do processamento do milho doce como volumoso. Os animais foram divididos em cinco grupos, totalizando 30 animais por tratamento: 1) relação volumoso: concentrado (V:C) de 50:50 do início ao final do experimento; 2) 30 dias com 100% V, seguidos de 30 dias com relação V:C de 50:50, e posteriormente relação V:C de 30:70 até o final do período experimental; 3) 45 dias iniciais do confinamento com 100% de volumoso, e posteriormente relação V:C de 50:50 até o final do período experimental; 4) 30 dias iniciais do confinamento com 100% de volumoso, e posteriormente relação V:C de 50:50 até o final do período experimental; 5) 100% de volumoso. O fornecimento exclusivo de volumoso durante o período total do confinamento proporcionou aos animais os menores peso vivo final, ganho médio diário e consumo de matéria seca, porém, apresentaram a melhor conversão alimentar entre os tratamentos avaliados e menor custo por arroba produzida. A inclusão de concentrado proporcionou, de maneira geral, maior desempenho dos animais, porém, aumento dos custos de produção.(AU)

Feedlot cattle performance was evaluated upon diets offering different levels and frequencies of sweet corn residue as source of roughage. The animals were divided into five groups, in a total of 30 animals per treatment. 1) roughage:grain ratio (R:G) of 50:50 from the start to the end of experiment , 2) 30 days with 100% roughage, followed by 30 days with R:G ratio of 50:50, and then R:G ratio of 30:70 until the end of the experimental period; 3) first 45 days at feedlot condition with 100% of roughage and then R:G ratio of 50:50 until the end of experimental period; 4) 30 days at start of feedlot with 100% of roughage, and then a ratio of 50:50 until the end of experimental period; 5) 100% of roughage. The lowest body weight, average daily gain and dry matter intake were achieved by animals receiving only roughage during the total period of feedlot; however, these animals presented the best feed conversion between evaluated treatments and lowest cost per pound produced. The concentrate inclusion offered, in general, highest animal performance, but with high-cost production.(AU)

Serological occurrence for tick-borne agents in beef cattle in the Brazilian Pantanal / Ocorrência sorológica para agentes transmitidos por carrapatos em bovinos de corte no Pantanal Brasileiro

This study investigated the seropositivity for five different tick-borne agents, namely Anaplasma marginale, Babesia bovis, Babesia bigemina, Coxiella burnetii, Anaplasma phagocytophilum, and Trypanosoma vivax in beef cattle in the Brazilian Pantanal. The serum samples collected from animals (200 cows; 200 calves) were used in indirect enzyme-linked immunosorbent assays (iELISA) to detect IgG antibodies against A. marginale, B. bovis, B. bigemina, and T. vivax, and Indirect Fluorescent Antibody Test (IFAT) for detecting IgG antibodies against C. burnetii and A. phagocytophilum. No correlation was observed between seropositivity for C. burnetii and A. phagocytophilum with other agents whereas moderate correlation was observed for A. marginalexB. bigemina x B. bovis. Cows were more seropositive for T. vivax whereas calves were more seropositive for B. bovis and B. bigemina. The highest number of seropositive animals by a single agent was observed for T. vivax (15.2%). Co-seropositivity for T. vivax + A. marginale was higher in cows (25.5%) and for T. vivax + B. bovis + B. bigemina + A. marginale was higher in calves (57.5%). The high seropositivity correlation for A. marginale x B. bovis x B. bigemina is probably due to the presence of the tick biological vector, Rhipicephalus microplus, in the studied farms. Common transmission pathways, mediated by hematophagous dipterans and fomites, may explain the high co-seropositivity of cows for A. marginale and T. vivax. Low seropositivity to C. burnetii is probably due to the type of breeding system employed (extensive). Seropositivity for A. phagocytophilum in only one animal suggests the occurrence of a cross-serological reaction with another agent of the genus Anaplasma.(AU)

Este estudo teve como objetivo determinar a co-soropositividade para agentes transmitidos por carrapatos, como Anaplasma marginale, Babesia bovis, Babesia bigemina, Coxiella burnetii, Anaplasma phagocytophilum, e Trypanosoma vivax em bovinos de corte do Pantanal Brasileiro. Amostras de soro foram colhidas de 400 animais (200 vacas; 200 bezerros) e submetidas a Ensaios Imunoenzimáticos Indiretos (iELISA) para detecção de anticorpos IgG anti- A. marginale, anti- B. bovis, anti- B. bigemina e anti- T. vivax, e à Reação de Imunofluorescência Indireta (RIFI) para detecção de anticorpos IgG anti -C. burnetii e anti- A. phagocytophilum. Ausência de correlação foi vista entre os animais soropositivos para C. burnetii e A. phagocytophilum com os outros agentes e correlação moderada ocorreu entre A. marginale x B. bigemina x B. bovis. Vacas foram mais soropositivas que bezerros para T. vivax e bezerros mais soropositivos que vacas para B. bovis e B. bigemina. Maior número de animais soropositivos para um único agente foi visto para T. vivax (15,2%). Vacas demonstraram maior co-soropositividade para T. vivax + A. marginale (25,5%) e bezerros para T. vivax + B. bovis + B. bigemina + A. marginale (57,5%). A alta correlação entre a soropositividade para A. marginale x B. bovis x B. bigemina é provavelmente devida à presença do vetor biológico, o carrapato Rhipicephalus microplus, nas fazendas estudadas. As vias de transmissão comuns, mediadas por dípteros hematófagos e fômites, podem explicar a alta co-soropositividade das vacas para A. marginale e T. vivax. A baixa soropositividade para C. burnetii é provavelmente devida ao tipo de sistema de criação empregado (extenso). A soropositividade para A. phagocytophilum em apenas um animal sugere a ocorrência de reação sorológica cruzada com outro agente do gênero Anaplasma.(AU)

Serological occurrence for tick-borne agents in beef cattle in the Brazilian Pantanal / Ocorrência sorológica para agentes transmitidos por carrapatos em bovinos de corte no Pantanal Brasileiro

This study investigated the seropositivity for five different tick-borne agents, namely Anaplasma marginale, Babesia bovis, Babesia bigemina, Coxiella burnetii, Anaplasma phagocytophilum, and Trypanosoma vivax in beef cattle in the Brazilian Pantanal. The serum samples collected from animals (200 cows; 200 calves) were used in indirect enzyme-linked immunosorbent assays (iELISA) to detect IgG antibodies against A. marginale, B. bovis, B. bigemina, and T. vivax, and Indirect Fluorescent Antibody Test (IFAT) for detecting IgG antibodies against C. burnetii and A. phagocytophilum. No correlation was observed between seropositivity for C. burnetii and A. phagocytophilum with other agents whereas moderate correlation was observed for A. marginalexB. bigemina x B. bovis. Cows were more seropositive for T. vivax whereas calves were more seropositive for B. bovis and B. bigemina. The highest number of seropositive animals by a single agent was observed for T. vivax (15.2%). Co-seropositivity for T. vivax + A. marginale was higher in cows (25.5%) and for T. vivax + B. bovis + B. bigemina + A. marginale was higher in calves (57.5%). The high seropositivity correlation for A. marginale x B. bovis x B. bigemina is probably due to the presence of the tick biological vector, Rhipicephalus microplus, in the studied farms. Common transmission pathways, mediated by hematophagous dipterans and fomites, may explain the high co-seropositivity of cows for A. marginale and T. vivax. Low seropositivity to C. burnetii is probably due to the type of breeding system employed (extensive). Seropositivity for A. phagocytophilum in only one animal suggests the occurrence of a cross-serological reaction with another agent of the genus Anaplasma.(AU)

Este estudo teve como objetivo determinar a co-soropositividade para agentes transmitidos por carrapatos, como Anaplasma marginale, Babesia bovis, Babesia bigemina, Coxiella burnetii, Anaplasma phagocytophilum, e Trypanosoma vivax em bovinos de corte do Pantanal Brasileiro. Amostras de soro foram colhidas de 400 animais (200 vacas; 200 bezerros) e submetidas a Ensaios Imunoenzimáticos Indiretos (iELISA) para detecção de anticorpos IgG anti- A. marginale, anti- B. bovis, anti- B. bigemina e anti- T. vivax, e à Reação de Imunofluorescência Indireta (RIFI) para detecção de anticorpos IgG anti -C. burnetii e anti- A. phagocytophilum. Ausência de correlação foi vista entre os animais soropositivos para C. burnetii e A. phagocytophilum com os outros agentes e correlação moderada ocorreu entre A. marginale x B. bigemina x B. bovis. Vacas foram mais soropositivas que bezerros para T. vivax e bezerros mais soropositivos que vacas para B. bovis e B. bigemina. Maior número de animais soropositivos para um único agente foi visto para T. vivax (15,2%). Vacas demonstraram maior co-soropositividade para T. vivax + A. marginale (25,5%) e bezerros para T. vivax + B. bovis + B. bigemina + A. marginale (57,5%). A alta correlação entre a soropositividade para A. marginale x B. bovis x B. bigemina é provavelmente devida à presença do vetor biológico, o carrapato Rhipicephalus microplus, nas fazendas estudadas. As vias de transmissão comuns, mediadas por dípteros hematófagos e fômites, podem explicar a alta co-soropositividade das vacas para A. marginale e T. vivax. A baixa soropositividade para C. burnetii é provavelmente devida ao tipo de sistema de criação empregado (extenso). A soropositividade para A. phagocytophilum em apenas um animal sugere a ocorrência de reação sorológica cruzada com outro agente do gênero Anaplasma.(AU)

Streamlined downstream process for efficient and sustainable (Fab)2 antivenom preparation

Antivenoms are the only validated treatment against snakebite envenoming. Numerous drawbacks pertaining to their availability, safety and efficacy are becoming increasingly evident due to low sustainability of current productions. Technological innovation of procedures generating therapeutics of higher purity and better physicochemical characteristics at acceptable cost is necessary. The objective was to develop at laboratory scale a compact, feasible and economically viable platform for preparation of equine F(ab')2 antivenom against Vipera ammodytes ammodytes venom and to support it with efficiency data, to enable estimation of the process cost-effectiveness. Methods: The principle of simultaneous caprylic acid precipitation and pepsin digestion has been implemented into plasma downstream processing. Balance between incomplete IgG breakdown, F(ab')2 over-digestion and loss of the active drug's protective efficacy was achieved by adjusting pepsin to a 1:30 substrate ratio (w/w) and setting pH at 3.2. Precipitation and digestion co-performance required 2 h-long incubation at 21 °C. Final polishing was accomplished by a combination of diafiltration and flow-through chromatography. In vivo neutralization potency of the F(ab')2 product against the venom's lethal toxicity was determined. Results: Only three consecutive steps, performed under finely tuned conditions, were sufficient for preservation of the highest process recovery with the overall yield of 74%, comparing favorably to others. At the same time, regulatory requirements were met. Final product was aggregate- and pepsin-free. Its composition profile was analyzed by mass spectrometry as a quality control check. Impurities, present in minor traces, were identified mostly as IgG/IgM fragments, contributing to active drug. Specific activity of the F(ab')2 preparation with respect to the plasma was increased 3.9-fold. Conclusion: A highly streamlined mode for production of equine F(ab')2 antivenom was engineered. In addition to preservation of the highest process yield and fulfillment of the regulatory demands, performance simplicity and rapidity in the laboratory setting were demonstrated. Suitability for large-scale manufacturing appears promising.(AU)

Streamlined downstream process for efficient and sustainable (Fab)2 antivenom preparation

Antivenoms are the only validated treatment against snakebite envenoming. Numerous drawbacks pertaining to their availability, safety and efficacy are becoming increasingly evident due to low sustainability of current productions. Technological innovation of procedures generating therapeutics of higher purity and better physicochemical characteristics at acceptable cost is necessary. The objective was to develop at laboratory scale a compact, feasible and economically viable platform for preparation of equine F(ab')2 antivenom against Vipera ammodytes ammodytes venom and to support it with efficiency data, to enable estimation of the process cost-effectiveness. Methods: The principle of simultaneous caprylic acid precipitation and pepsin digestion has been implemented into plasma downstream processing. Balance between incomplete IgG breakdown, F(ab')2 over-digestion and loss of the active drug's protective efficacy was achieved by adjusting pepsin to a 1:30 substrate ratio (w/w) and setting pH at 3.2. Precipitation and digestion co-performance required 2 h-long incubation at 21 °C. Final polishing was accomplished by a combination of diafiltration and flow-through chromatography. In vivo neutralization potency of the F(ab')2 product against the venom's lethal toxicity was determined. Results: Only three consecutive steps, performed under finely tuned conditions, were sufficient for preservation of the highest process recovery with the overall yield of 74%, comparing favorably to others. At the same time, regulatory requirements were met. Final product was aggregate- and pepsin-free. Its composition profile was analyzed by mass spectrometry as a quality control check. Impurities, present in minor traces, were identified mostly as IgG/IgM fragments, contributing to active drug. Specific activity of the F(ab')2 preparation with respect to the plasma was increased 3.9-fold. Conclusion: A highly streamlined mode for production of equine F(ab')2 antivenom was engineered. In addition to preservation of the highest process yield and fulfillment of the regulatory demands, performance simplicity and rapidity in the laboratory setting were demonstrated. Suitability for large-scale manufacturing appears promising.(AU)

Maturação in vitro de oócitos de cutias (Dasyprocta prymnolopha, Wagler 1831) seguida de fertilização in vitro e ativação partenogenética / In vitro maturation of agoutis (Dasyprocta prymnolopha) oocytes followed by in vitro fertilization and parthenogenetic activation

O objetivo foi avaliar protocolos de maturação in vitro (MIV) para oócitos de cutias, seguida de fertilização in vitro (FIV) e ativação partenogenética (AP). Os oócitos imaturos (CCOs) foram obtidos por fatiamento do ovário, após OSH, e submetidos a três grupos: MAT - 16 (16 horas de maturação), MAT - 20 (20 horas de maturação) e MAT - 24 (24 horas de maturação), em incubadora de cultivo a 38,8°C, com atmosfera de 5% de CO2 e 95% de umidade relativa. A maturação foi analisada pela presença do primeiro corpúsculo polar. Em seguida, os CCOs maduros foram submetidos à FIV, com período de coincubação dos CCOs e dos espermatozoides de 15h, a 38,8ºC e 5% de CO2, e AP com ionomicina. Os grupos de MIV foram analisados utilizando-se o teste qui-quadrado e, nos experimentos de FIV e AP, foram analisadas a taxa de clivagem e a proporção de desenvolvimento embrionário. A análise estatística foi realizada utilizando-se o programa SAS. Houve diferença significativa entre os grupos de maturação, tendo os grupos MAT - 20 e MAT - 24 apresentado maior porcentagem de oócitos maturados in vitro. As taxas de clivagem e de desenvolvimento embrionário foram de 8,6% e 2,9%, respectivamente, na FIV, e de 63,6% e 15,1%, na AP. Entretanto, nos dois casos, o embrião não passou do estágio de mórula.(AU)

The objective was to evaluate IVM protocols for agouti oocytes, followed by in vitro fertilization (IVF) and parthenogenetic activation (PA). The immature oocytes (CCOs) were obtained by slicing the ovary after OSH and submitted to three groups: MAT - 16 (16 hours maturation), MAT - 20 (20 hours maturation) and MAT - (24 hours maturation), in a culture incubator at 38.8°C, with an atmosphere of 5% CO2 and 95% relative humidity. The maturation was analyzed by the presence of the first polar corpuscle. Then, mature CCOs were submitted to IVF, with co-incubation period of CCOs and spermatozoa from 15h to 38.8°C and 5% of CO2, and PA with inomycin. The IVM groups were analyzed using the chi-square test and in the FIV and PA experiment the rate of cleavage and the rate of embryonic development were analyzed. Statistical analysis was performed using the SAS program. There was a significant difference between the maturation groups, and the MAT - 20 and MAT - 24 groups showed a higher percentage of matured oocytes in vitro. The rates of cleavage and embryonic development were 8.6% and 2.9%, respectively in FIV and 63.6% and 15.1% in PA. However, in both cases the embryo did not pass beyond the morula stage.(AU)

Maturação in vitro de oócitos de cutias (Dasyprocta prymnolopha, Wagler 1831) seguida de fertilização in vitro e ativação partenogenética / In vitro maturation of agoutis (Dasyprocta prymnolopha) oocytes followed by in vitro fertilization and parthenogenetic activation

O objetivo foi avaliar protocolos de maturação in vitro (MIV) para oócitos de cutias, seguida de fertilização in vitro (FIV) e ativação partenogenética (AP). Os oócitos imaturos (CCOs) foram obtidos por fatiamento do ovário, após OSH, e submetidos a três grupos: MAT - 16 (16 horas de maturação), MAT - 20 (20 horas de maturação) e MAT - 24 (24 horas de maturação), em incubadora de cultivo a 38,8°C, com atmosfera de 5% de CO2 e 95% de umidade relativa. A maturação foi analisada pela presença do primeiro corpúsculo polar. Em seguida, os CCOs maduros foram submetidos à FIV, com período de coincubação dos CCOs e dos espermatozoides de 15h, a 38,8ºC e 5% de CO2, e AP com ionomicina. Os grupos de MIV foram analisados utilizando-se o teste qui-quadrado e, nos experimentos de FIV e AP, foram analisadas a taxa de clivagem e a proporção de desenvolvimento embrionário. A análise estatística foi realizada utilizando-se o programa SAS. Houve diferença significativa entre os grupos de maturação, tendo os grupos MAT - 20 e MAT - 24 apresentado maior porcentagem de oócitos maturados in vitro. As taxas de clivagem e de desenvolvimento embrionário foram de 8,6% e 2,9%, respectivamente, na FIV, e de 63,6% e 15,1%, na AP. Entretanto, nos dois casos, o embrião não passou do estágio de mórula.(AU)

The objective was to evaluate IVM protocols for agouti oocytes, followed by in vitro fertilization (IVF) and parthenogenetic activation (PA). The immature oocytes (CCOs) were obtained by slicing the ovary after OSH and submitted to three groups: MAT - 16 (16 hours maturation), MAT - 20 (20 hours maturation) and MAT - (24 hours maturation), in a culture incubator at 38.8°C, with an atmosphere of 5% CO2 and 95% relative humidity. The maturation was analyzed by the presence of the first polar corpuscle. Then, mature CCOs were submitted to IVF, with co-incubation period of CCOs and spermatozoa from 15h to 38.8°C and 5% of CO2, and PA with inomycin. The IVM groups were analyzed using the chi-square test and in the FIV and PA experiment the rate of cleavage and the rate of embryonic development were analyzed. Statistical analysis was performed using the SAS program. There was a significant difference between the maturation groups, and the MAT - 20 and MAT - 24 groups showed a higher percentage of matured oocytes in vitro. The rates of cleavage and embryonic development were 8.6% and 2.9%, respectively in FIV and 63.6% and 15.1% in PA. However, in both cases the embryo did not pass beyond the morula stage.(AU)