Efficacy of lapachol on treatment of cutaneous and visceral leishmaniasis.

Leishmaniasis is one of the most important neglected diseases worldwide. It is a life-threatening disease and causes significant morbidity, long-term disability, and early death. Treatment involves disease control or use of intervention measures, although the currently used drugs require long-lasting therapy, and display toxicity and reduced efficacy. The use of natural products isolated from plants, such as lapachol, an abundant naphthoquinone naturally occurring in South American Handroanthus species (Tabebuia, Bignoniaceae), is a promising option for the treatment of leishmaniasis. In this study, we investigated the leishmanicidal activity of lapachol in vitro and in vivo against Leishmania infantum and L. amazonensis, causative agents of visceral and cutaneous leishmaniasis, respectively. Low cytotoxicity in HepG2 cells (3405.8 ±â€¯261.33 µM), good anti-Leishmania activity, and favorable selectivity indexes (SI) against promastigotes of both L. amazonensis (IC50 = 79.84 ±â€¯9.10 µM, SI = 42.65) and L. infantum (IC50 = 135.79 ±â€¯33.04 µM, SI = 25.08) were observed. Furthermore, anti-Leishmania activity assays performed on intracellular amastigotes showed good activity for lapachol (IC50 = 191.95 µM for L. amazonensis and 171.26 µM for L. infantum). Flow cytometric analysis demonstrated that the cytotoxic effect of lapachol in Leishmania promastigotes was caused by apoptosis-like death. Interestingly, the in vitro leishmanicidal effect of lapachol was confirmed in vivo in murine models of visceral and cutaneous leishmaniasis, as lapachol (25 mg/kg oral route for 24 h over 10 days) was able to significantly reduce the parasitic load in skin lesions, liver, and spleen, similar to amphotericin B, the reference drug. These results reinforce the therapeutic potential of lapachol, which warrants further investigations as an anti-leishmaniasis therapeutic.

Canine Leishmaniasis: An Overview of the Current Status and Strategies for Control.

Canine leishmaniasis (CanL) is a vector-borne disease caused by Leishmania infantum and is transmitted by female phlebotomine sand flies primarily between animals and secondarily to humans. The course of infection may be different from one individual dog to another, ranging from spontaneous cure to acute evolution that leads to death, if proper management and therapy are not adopted. A parasitological cure is rarely achieved and clinical recurrences in CanL are frequent. Vaccination associated with the use of topical insecticides is undoubtedly the most effective form of prevention and control of the disease. In order to integrate the most important scientific knowledge of the literature in one objective publication, this review proposes a short overview of the main points of CanL.

Effectiveness of Cratylia argentea as an animal feed supplement in the control of gastrointestinal nematodes in sheep / Eficácia de Cratylia argentea como suplemento alimentar animal no controle das nematodioses gastrointestinais em ovinos

Gastrointestinal nematodes are considered one of the most important sanitary problems affectingruminants globally; whether parasitizing alone or acting with other pathogens, they are responsiblefor severe economic losses related to animal husbandry. Endoparasites in small ruminants areusually controlled with anti-helminthic drugs. However, certain strains of endoparasites are resistantto such drugs. Supplementation of the diet with protein-rich foods is one strategy for reducing thenegative impact of gastrointestinal parasites. Cratylia argentea has anthelmintic properties and othercharacteristics that favor its use as an animal feed supplement. In the present study, we subdividedcrossbred lambs into homogeneous groups and fed them C. argentea (G1), C. argentea plus Brachiariaspp. (G2), or Brachiaria spp. (G3) only. The animals in group G1 had a lower average number of eggsper gram of feces (EPG) at Day 42 (p 0.05), indicating the absenceof anemia and normal globular volume. This reinforces the importance of the measurement of EPG; thisis a quantitative method and does not depend on the subjective interpretation of the results.

Os nematódeos gastrointestinais são considerados um dos problemas sanitários mais importantes dosruminantes no mundo, seja em infecções isoladas ou atuando com outros patógenos, são responsáveis porseveras perdas econômicas relacionadas à produção animal. O controle dos endoparasitas em pequenosruminantes normalmente é feito por meio de drogas anti-helmínticas. No entanto, determinadas cepasde endoparasitos são resistentes a esses fármacos. A suplementação da dieta com alimentos ricos emproteína é uma estratégia para reduzir o impacto negativo das parasitoses gastrointestinais. Cratylia argentea tem efeito anti-helmíntico, além de outras características que favorecem seu uso comosuplemento para alimentação animal. No presente estudo, borregas cruzadas foram subdivididas emgrupos homogêneos e alimentadas com C. argentea (G1), C. argentea mais Brachiaria spp. (G2) ouapenas Brachiaria spp. (G3). Os animais do grupo G1 apresentaram menor valor médio de ovos porgrama de fezes (OPG) aos 42 dias (p0,05), indicando ausência de anemia e volume globularnormal. Isso reforça a importância da avaliação por OPG, que é um método quantitativo e não dependede interpretação subjetiva de resultados.

Effectiveness of Cratylia argentea as an animal feed supplement in the control of gastrointestinal nematodes in sheep / Eficácia de Cratylia argentea como suplemento alimentar animal no controle das nematodioses gastrointestinais em ovinos

Gastrointestinal nematodes are considered one of the most important sanitary problems affectingruminants globally; whether parasitizing alone or acting with other pathogens, they are responsiblefor severe economic losses related to animal husbandry. Endoparasites in small ruminants areusually controlled with anti-helminthic drugs. However, certain strains of endoparasites are resistantto such drugs. Supplementation of the diet with protein-rich foods is one strategy for reducing thenegative impact of gastrointestinal parasites. Cratylia argentea has anthelmintic properties and othercharacteristics that favor its use as an animal feed supplement. In the present study, we subdividedcrossbred lambs into homogeneous groups and fed them C. argentea (G1), C. argentea plus Brachiariaspp. (G2), or Brachiaria spp. (G3) only. The animals in group G1 had a lower average number of eggsper gram of feces (EPG) at Day 42 (p < 0.05), and the average count was ten times lower than that inthe control at the last evaluated time-point (Day 56). The animals exhibited linear weight gain, andthose in group G1 gained the most weight. The combined use of drugs and plants containing bioactivecompounds can minimize the evolution of multidrug resistance, prolonging the efficacy of the drugs.The FAMACHA® score showed little variation during the experiment (p > 0.05), indicating the absenceof anemia and normal globular volume. This reinforces the importance of the measurement of EPG; thisis a quantitative method and does not depend on the subjective interpretation of the results.(AU)

Os nematódeos gastrointestinais são considerados um dos problemas sanitários mais importantes dosruminantes no mundo, seja em infecções isoladas ou atuando com outros patógenos, são responsáveis porseveras perdas econômicas relacionadas à produção animal. O controle dos endoparasitas em pequenosruminantes normalmente é feito por meio de drogas anti-helmínticas. No entanto, determinadas cepasde endoparasitos são resistentes a esses fármacos. A suplementação da dieta com alimentos ricos emproteína é uma estratégia para reduzir o impacto negativo das parasitoses gastrointestinais. Cratylia argentea tem efeito anti-helmíntico, além de outras características que favorecem seu uso comosuplemento para alimentação animal. No presente estudo, borregas cruzadas foram subdivididas emgrupos homogêneos e alimentadas com C. argentea (G1), C. argentea mais Brachiaria spp. (G2) ouapenas Brachiaria spp. (G3). Os animais do grupo G1 apresentaram menor valor médio de ovos porgrama de fezes (OPG) aos 42 dias (p<0,05) e sua contagem média foi dez vezes mais baixa que a docontrole no último período de tempo avaliado (56 dias). Os animais apresentaram aumento linear doganho de peso e aqueles do grupo G1 apresentaram o maior ganho. O escore FAMACHA® mostroupouca variação durante o experimento (p>0,05), indicando ausência de anemia e volume globularnormal. Isso reforça a importância da avaliação por OPG, que é um método quantitativo e não dependede interpretação subjetiva de resultados.(AU)

Anthelmintic efficacy of Cratylia argentea (Desv. ) Kuntze against the gastrointestinal nematodes of sheep / Eficácia anti-helmíntica de Cratylia argentea (Desv. ) Kuntze contra nematódeos gastrintestinais de ovinos

The use of natural products having therapeutic properties can be considered an alternative approach for the control of helminths in ruminants, reducing the indiscriminate use of chemicals as well as production costs. The anthelmintic effectiveness of Cratylia argentea on sheep endoparasites was evaluated both in vitro by assessing the number of third-stage larvae recovered per gram of feces (LPG) from coprocultures treated with pods/ground seeds, macerated leaves or vermiculite (control), and in vivo using the Fecal Egg Count Reduction Test (FECRT) in two groups of lambs (n = 8/group) that received 1 mL kg-1 body weight of C. argentea solution (treated group) or distilled water (control group) orally. When compared to control group, there was a significant reduction in the percentage of LPG in the leaves group (55.88%; p ? 0.05) and pods/ground seeds group (82.48%; p ? 0.01). There was no significant reduction (p > 0.05) in the average count of EPG upon treatment with the aqueous solution of C. argentea. These findings suggest that C. argentea extract mainly obtained from pods/ground seeds has a promising potential for the control of young stages of gastrointestinal nematodes. However, further studies are needed to validate the use of C. argentea as an alternative method for the control of parasitic diseases of ruminants.(AU)

O uso de produtos naturais com propriedades terapêuticas pode ser considerado uma abordagem alternativa no controle de helmintos em ruminantes, reduzindo o uso indiscriminado de produtos químicos, bem como os custos de produção. A eficácia anti-helmíntica de Cratylia argentea sobre endoparasitos de ovinos foi avaliada tanto in vitro, mediante teste de recuperação de larvas infectantes por grama de fezes (LPG) a partir de coproculturas tratadas com vagens/sementes trituradas, folhas maceradas e vermiculita (controle), como in vivo, por meio de Teste de Redução da Contagem de Ovos por Grama de Fezes (TRCOF) em dois grupos de borregas (n = 8/grupo) que receberem por via oral 1 mL kg-1 PV de solução de C. argentea (grupo tratado) ou água destilada (grupo controle). Quando comparado ao grupo controle, houve redução estatisticamente significativa na porcentagem de LPG a partir do grupo tratado com folhas (55,88%; p ? 0,05) e do grupo tratado com vagens/sementes (82,48%; p ? 0,01). Não houve redução significativa (p > 0,05) na contagem média de OPG após o tratamento com a solução aquosa de C. argentea. Essas descobertas sugerem que o extrato de C. argentea obtido principalmente a partir de vagens/sementes tem potencial promissor para o controle de estágios jovens de nematódeos gastrointestinais. Contudo, são necessários novos estudos, a fim de validar o uso desta planta como um método alternativo no controle das parasitoses de ruminantes.(AU)

Anthelmintic efficacy of Cratylia argentea (Desv. ) Kuntze against the gastrointestinal nematodes of sheep / Eficácia anti-helmíntica de Cratylia argentea (Desv. ) Kuntze contra nematódeos gastrintestinais de ovinos

The use of natural products having therapeutic properties can be considered an alternative approach for the control of helminths in ruminants, reducing the indiscriminate use of chemicals as well as production costs. The anthelmintic effectiveness of Cratylia argentea on sheep endoparasites was evaluated both in vitro by assessing the number of third-stage larvae recovered per gram of feces (LPG) from coprocultures treated with pods/ground seeds, macerated leaves or vermiculite (control), and in vivo using the Fecal Egg Count Reduction Test (FECRT) in two groups of lambs (n = 8/group) that received 1 mL kg-1 body weight of C. argentea solution (treated group) or distilled water (control group) orally. When compared to control group, there was a significant reduction in the percentage of LPG in the leaves group (55.88%; p ? 0.05) and pods/ground seeds group (82.48%; p ? 0.01). There was no significant reduction (p > 0.05) in the average count of EPG upon treatment with the aqueous solution of C. argentea. These findings suggest that C. argentea extract mainly obtained from pods/ground seeds has a promising potential for the control of young stages of gastrointestinal nematodes. However, further studies are needed to validate the use of C. argentea as an alternative method for the control of parasitic diseases of ruminants.

O uso de produtos naturais com propriedades terapêuticas pode ser considerado uma abordagem alternativa no controle de helmintos em ruminantes, reduzindo o uso indiscriminado de produtos químicos, bem como os custos de produção. A eficácia anti-helmíntica de Cratylia argentea sobre endoparasitos de ovinos foi avaliada tanto in vitro, mediante teste de recuperação de larvas infectantes por grama de fezes (LPG) a partir de coproculturas tratadas com vagens/sementes trituradas, folhas maceradas e vermiculita (controle), como in vivo, por meio de Teste de Redução da Contagem de Ovos por Grama de Fezes (TRCOF) em dois grupos de borregas (n = 8/grupo) que receberem por via oral 1 mL kg-1 PV de solução de C. argentea (grupo tratado) ou água destilada (grupo controle). Quando comparado ao grupo controle, houve redução estatisticamente significativa na porcentagem de LPG a partir do grupo tratado com folhas (55,88%; p ? 0,05) e do grupo tratado com vagens/sementes (82,48%; p ? 0,01). Não houve redução significativa (p > 0,05) na contagem média de OPG após o tratamento com a solução aquosa de C. argentea. Essas descobertas sugerem que o extrato de C. argentea obtido principalmente a partir de vagens/sementes tem potencial promissor para o controle de estágios jovens de nematódeos gastrointestinais. Contudo, são necessários novos estudos, a fim de validar o uso desta planta como um método alternativo no controle das parasitoses de ruminantes.

Relationship between clinical and pathological signs and severity of canine leishmaniasis.

Canine visceral leishmaniasis (CVL) is a zoonotic disease that presents variable clinical and laboratory aspects. The aims of this study were to identify the main biochemical/hematological status of dogs naturally infected with Leishmania (Leishmania) infantum and to associate theses parameters with clinical forms of CVL. Blood samples were analyzed from 51 dogs, 15 uninfected (control group) and 36 infected, which were classified clinically in three groups: asymptomatic (n=12), oligosymptomatic (n=12) and symptomatic (n=12). All the infected dogs showed lower albumin/globulin ratios (A-G ratio) than the limit of reference. The mean values of total protein, urea, α-globulin 2, globulin and A-G ratio of infected dogs were outside the reference interval and differed significantly from those of the controls. Anemia was detected only in groups that showed clinical signs of the disease, and a statistical analysis indicated a significantly higher frequency of lower eritrogram in these groups than in the asymptomatic group. In addition, a significant association was observed between anemia and the presence of the symptoms, with dogs displaying higher erythrogram values showing better clinical conditions. These results provide additional evidence that the clinical forms of CVL may reflect on the erythrogram status.

Relationship between clinical and pathological signs and severity of canine leishmaniasis / Relação entre sinais clínico-patológicos e gravidade da leishmaniose visceral canina

Canine visceral leishmaniasis (CVL) is a zoonotic disease that presents variable clinical and laboratory aspects. The aims of this study were to identify the main biochemical/hematological status of dogs naturally infected with Leishmania (Leishmania) infantum and to associate theses parameters with clinical forms of CVL. Blood samples were analyzed from 51 dogs, 15 uninfected (control group) and 36 infected, which were classified clinically in three groups: asymptomatic (n=12), oligosymptomatic (n=12) and symptomatic (n=12). All the infected dogs showed lower albumin/globulin ratios (A-G ratio) than the limit of reference. The mean values of total protein, urea, α-globulin 2, globulin and A-G ratio of infected dogs were outside the reference interval and differed significantly from those of the controls. Anemia was detected only in groups that showed clinical signs of the disease, and a statistical analysis indicated a significantly higher frequency of lower eritrogram in these groups than in the asymptomatic group. In addition, a significant association was observed between anemia and the presence of the symptoms, with dogs displaying higher erythrogram values showing better clinical conditions. These results provide additional evidence that the clinical forms of CVL may reflect on the erythrogram status.

A leishmaniose visceral canina (LVC) é uma zoonose com aspectos clínicos e laboratoriais variáveis. O objetivo deste trabalho foi identificar os principais achados hematológicos e bioquímicos em cães naturalmente infectados com Leishmania (Leishmania) infantum e associar esses parâmetros com as formas clínicas da LVC. Foram analisadas amostras sanguíneas provenientes de 51 cães, sendo 15 cães não infectados (grupo controle) e 36 infectados, os quais foram classificados clinicamente em três grupos: assintomáticos (n=12), oligossintomáticos (n=12) e sintomáticos (n=12). Todos os cães infectados apresentaram valores na relação albumina/globulina (A/G) abaixo do limite inferior de referência. Os valores médios de proteína total, uréia, α-2 globulina, globulina e A/G dos grupos de cães infectados permaneceram fora dos intervalos de referências e significativamente diferente quando comparados aos do grupo controle. Anemia foi registrada somente nos grupos de animais que manifestavam sinais clínicos da enfermidade, sendo que nas análises estatísticas constatou-se frequência significativamente maior de alterações no eritrograma quando comparados ao grupo assintomático. Associação significativa foi observada entre anemia e a presença de sinais clínicos, onde os cães com os maiores valores de eritrograma apresentavam a melhor condição clínica. Os resultados fornecem evidência adicional que as formas clínicas da LVC podem refletir no eritrograma.

Parasites of domestic and wild canids in the region of Serra do Cipó National Park, Brazil.

Over recent decades, diseases have been shown to be important causes of extinctions among wild species. Greater emphasis has been given to diseases transmitted by domestic animals, which have been increasing in numbers in natural areas, along with human populations. This study had the aim of investigating the presence of intestinal helminths in wild canids (maned wolf, Chrysocyon brachyurus, and crab-eating fox, Cerdocyon thous) in the Serra do Cipó National Park (43-44º W and 19-20º S) and endo and ectoparasites of domestic dogs in the Morro da Pedreira Environmental Protection Area (an area surrounding the National Park). The Serra do Cipó is located in the state of Minas Gerais, Brazil. Among the enteroparasites found in domestic and wild canids, the following taxons were identified: Ancylostomidae, Trichuridae, Toxocara sp., Spirocerca sp., Physaloptera sp., Strongyloides sp., Cestoda, Dipylidium caninum, Diphyllobothriidae, Hymenolepidae, Anoplocephalidae, Trematoda, Acanthocephala and Isospora sp. Domestic dogs were positive for leishmaniasis and Babesia canis in serological tests. Among the ectoparasites, Rhipicephalus sanguineus, Amblyomma cajennense and Ctenocephalides felis felis were observed in domestic dogs. Variations in the chaetotaxy of the meta-episternum and posterior tibia were observed in some specimens of C. felis felis.

Parasites of domestic and wild canids in the region of Serra do Cipó National Park, Brazil / Parasitos de canídeos domésticos e silvestres na região do Parque Nacional da Serra do Cipó - Brasil

Over recent decades, diseases have been shown to be important causes of extinctions among wild species. Greater emphasis has been given to diseases transmitted by domestic animals, which have been increasing in numbers in natural areas, along with human populations. This study had the aim of investigating the presence of intestinal helminths in wild canids (maned wolf, Chrysocyon brachyurus, and crab-eating fox, Cerdocyon thous) in the Serra do Cipó National Park (43-44º W and 19-20º S) and endo and ectoparasites of domestic dogs in the Morro da Pedreira Environmental Protection Area (an area surrounding the National Park). The Serra do Cipó is located in the state of Minas Gerais, Brazil. Among the enteroparasites found in domestic and wild canids, the following taxons were identified: Ancylostomidae, Trichuridae, Toxocara sp., Spirocerca sp., Physaloptera sp., Strongyloides sp., Cestoda, Dipylidium caninum, Diphyllobothriidae, Hymenolepidae, Anoplocephalidae, Trematoda, Acanthocephala and Isospora sp. Domestic dogs were positive for leishmaniasis and Babesia canis in serological tests. Among the ectoparasites, Rhipicephalus sanguineus, Amblyomma cajennense and Ctenocephalides felis felis were observed in domestic dogs. Variations in the chaetotaxy of the meta-episternum and posterior tibia were observed in some specimens of C. felis felis.

Nas últimas décadas, as doenças têm sido apontadas como importantes causas de extinção de espécies silvestres. Maior ênfase tem sido dada às doenças transmitidas por animais domésticos que crescem em número, bem como as populações humanas, em áreas naturais. O presente estudo objetivou verificar a presença de helmintos intestinais de canídeos silvestres (lobo-guará - Chrysocyon brachyurus e cachorro-do-mato - Cerdocyon thous) do Parque Nacional da Serra do Cipó (43-44º W e 19-20º S) e endo e ectoparasitos de cães domésticos da Área de Proteção Ambiental Morro da Pedreira (entorno do Parque Nacional). A Serra do Cipó está localizada no Estado de Minas Gerais, Brasil. Entre os enteroparasitos encontrados em canídeos domésticos e silvestres, os táxons seguintes, Ancylostomidae, Trichuridae, Toxocara sp., Spirocerca sp., Physaloptera sp., Strongyloides sp., Cestoda, Dipylidium caninum, Diphyllobothriidae, Hymenolepidae, Anoplocephalidae, Trematoda, Acanthocephala e Isospora sp. foram identificados. Cães domésticos foram positivos para leishmaniose e Babesia canis nos testes sorológicos. Entre os ectoparasitos, Rhipicephalus sanguineus, Amblyomma cajennense, Ctenocephalides felis felis foram observados em cães domésticos. Variações na quetotaxia do metepisterno e tíbia posterior foram observados em alguns espécimes de C. felis felis.

Parasites of domestic and wild canids in the region of Serra do Cipó National Park, Brazil / Parasitos de canídeos domésticos e silvestres na região do Parque Nacional da Serra do Cipó - Brasil

Over recent decades, diseases have been shown to be important causes of extinctions among wild species. Greater emphasis has been given to diseases transmitted by domestic animals, which have been increasing in numbers in natural areas, along with human populations. This study had the aim of investigating the presence of intestinal helminths in wild canids (maned wolf, Chrysocyon brachyurus, and crab-eating fox, Cerdocyon thous) in the Serra do Cipó National Park (43-44º W and 19-20º S) and endo and ectoparasites of domestic dogs in the Morro da Pedreira Environmental Protection Area (an area surrounding the National Park). The Serra do Cipó is located in the state of Minas Gerais, Brazil. Among the enteroparasites found in domestic and wild canids, the following taxons were identified: Ancylostomidae, Trichuridae, Toxocara sp., Spirocerca sp., Physaloptera sp., Strongyloides sp., Cestoda, Dipylidium caninum, Diphyllobothriidae, Hymenolepidae, Anoplocephalidae, Trematoda, Acanthocephala and Isospora sp. Domestic dogs were positive for leishmaniasis and Babesia canis in serological tests. Among the ectoparasites, Rhipicephalus sanguineus, Amblyomma cajennense and Ctenocephalides felis felis were observed in domestic dogs. Variations in the chaetotaxy of the meta-episternum and posterior tibia were observed in some specimens of C. felis felis.(AU)

Nas últimas décadas, as doenças têm sido apontadas como importantes causas de extinção de espécies silvestres. Maior ênfase tem sido dada às doenças transmitidas por animais domésticos que crescem em número, bem como as populações humanas, em áreas naturais. O presente estudo objetivou verificar a presença de helmintos intestinais de canídeos silvestres (lobo-guará - Chrysocyon brachyurus e cachorro-do-mato - Cerdocyon thous) do Parque Nacional da Serra do Cipó (43-44º W e 19-20º S) e endo e ectoparasitos de cães domésticos da Área de Proteção Ambiental Morro da Pedreira (entorno do Parque Nacional). A Serra do Cipó está localizada no Estado de Minas Gerais, Brasil. Entre os enteroparasitos encontrados em canídeos domésticos e silvestres, os táxons seguintes, Ancylostomidae, Trichuridae, Toxocara sp., Spirocerca sp., Physaloptera sp., Strongyloides sp., Cestoda, Dipylidium caninum, Diphyllobothriidae, Hymenolepidae, Anoplocephalidae, Trematoda, Acanthocephala e Isospora sp. foram identificados. Cães domésticos foram positivos para leishmaniose e Babesia canis nos testes sorológicos. Entre os ectoparasitos, Rhipicephalus sanguineus, Amblyomma cajennense, Ctenocephalides felis felis foram observados em cães domésticos. Variações na quetotaxia do metepisterno e tíbia posterior foram observados em alguns espécimes de C. felis felis.(AU)

New mucoadhesive chitosan film for ophthalmic drug delivery of timolol maleate: in vivo evaluation.

PURPOSE: Chitosan, a cationic polysaccharide biopolymer with mucoadhesive properties, presents a promising future in the prolonged ocular delivery of drugs. The present study compared the efficacy and safety of chitosan-coated timolol maleate (TM) mucoadhesive film, using a 0.5% TM commercial ophthalmic solution in a rabbit model. In addition, this study investigates the maximum release time of these implants in vivo. METHODS: The mucoadhesive films were prepared by means of a casting and solvent evaporation technique performed in a 2 wt% acetic acid solution and distilled water. Physical properties were characterized by release and swelling studies, differential scanning calorimetry, and attenuated total reflectance fourier transformed infrared spectroscopy (ATR-FTIR). The developed formulations were evaluated for their pharmacodynamics in ocular normotensive albino rabbits, in which the intraocular pressure (IOP) was measured by means of applanation tonometer on alternative days (13 h) for 11 weeks. For 15 days, 0.5% TM commercial ophthalmic solution was administered twice a day (n=5) and compared to chitosan-coated TM (n=5). In the control group (n=5), saline was used twice a day. The maximum TM release time from chitosan films were also recorded. After euthanasia, the right eyes were removed from the 3 groups for histological analyses. RESULTS: In an in vitro study, TM was released over a 4-week period, in which 85% of the drug was released over the first 2 weeks. However, the film's release of TM lowered the in vivo IOP levels over a 10-week period. No significant difference in the lowering of IOP in rabbits treated with 0.5% TM commercial ophthalmic solution, as compared to those that received the films (P<0.05), could be observed. No signs of ocular discomfort or irritations could be identified upon ophthalmic examination by slit-lamp biomicroscopy. Ophthalmic structures that came in direct contact with the films revealed no alterations within the histopathological studies. Moreover, the animals showed no signs of ocular discomfort during the experimental assays. CONCLUSION: These findings suggest that the TM-loaded chitosan film is safe and efficient and presents a promising future as an ocular drug delivery system in the treatment and prevention of glaucoma.

First report of infection of Lutzomyia longipalpis by Leishmania (Leishmania) infantum from a naturally infected cat of Brazil.

In recent years, cases of feline visceral leishmaniasis (FVL) have been described in different countries. In urban areas, domestic cats are suggested as possible alternative reservoirs of Leishmania (L.) infantum, the causal agent of visceral leishmaniasis (VL). This paper reports the first case of infection of Lutzomyia longipalpis by L. infantum of a naturally infected cat from Brazil through xenodiagnosis. The presence of a cat with FVL and its infectivity to the natural vector in Belo Horizonte city, an endemic area of VL in Brazil, suggests the need for further studies to determine the rate of occurrence of FVL among domestic cats and the infectivity ratio of L. longipalpis in endemic areas, and what is the role of these animals in the epidemiology of the disease.

Hepatic extracellular matrix alterations in dogs naturally infected with Leishmania (Leishmania) chagasi.

Summary The aim of this work was to study alterations in the extracellular matrix of liver in dogs naturally infected with Leishmania (Leishmania) chagasi that are correlated with clinical aspects and with histological, parasitological and immunological findings. The study was carried out on 30 dogs, 10 uninfected (control group) and 20 infected. The infected animals were further divided into two groups: an asymptomatic group of 10 dogs without clinical signs of the disease; and a symptomatic group of 10 dogs with classical clinical signs. All thirty animals were mongrel dogs of undefined age, obtained from the municipality of Belo Horizonte, MG, metropolitan area. During necropsy, liver fragments were collected and fixed in 10% buffered formaldehyde for histological examination. Paraffined sections of the tissues were stained with haematoxylin-eosin, Gomori's ammoniacal silver stain for reticular fibres and strepto-avidin peroxidase for immunohistochemical detection of Leishmania amastigotes. Frozen tissue sections were stained by immunofluorescence for fibronectin (FN) and laminin (LN). Liver collagen deposition was significantly greater in the infected than the control animals and differed significantly between the symptomatic and asymptomatic dogs. There was a positive correlation between the parasite load and liver collagen deposition. The increased collagen deposition in infected animal livers may be associated with the parasite burden. Adhesive FN and LN fibres were significantly more highly expressed in the livers of symptomatic than of asymptomatic dogs. Our results demonstrate that canine visceral leishmaniasis causes fibrogenesis in liver, associated with the parasite load and degenerative processes.

First report of vertical transmission of Leishmania (Leishmania) infantum in a naturally infected bitch from Brazil.

Dogs are the most important reservoir of Leishmania (L.) infantum, the causal agent of visceral leishmaniasis (VL) in Brazil. Vectorial infection is the main route of transmission of the parasites. This paper reports the first case of vertical transmission of L. infantum in Brazil, confirmed by PCR and immunohistochemistry techniques in samples from spleen and liver of two stillborn pups from a bitch naturally infected with L. infantum in Belo Horizonte city, endemic area of VL. This result confirms the existence of transplacental transmission of Leishmania between dogs, and suggests the need for further studies to determine the rate of occurrence of this fact in endemic areas and what is their role in the epidemiology of the disease.