In vitro effect of Chrysosporium indicum and Chrysosporium keratinophylum on Toxocara canis eggs.

The degree of antagonism exercised by fungi on geohelminth development varies according to the morphological alterations caused by different fungal species. Saprophytic fungi may exert ovicidal or ovistatic effects. The aim of this study was to apply scanning electron microscopy (SEM) to observe the action of two soil saprophytic species of Chrysosporium (C. indicum and C. keratinophylum) on Toxocara canis eggs. The fungal strains to be tested were incubated for 28 days at 28°C in 2% water agar with a suspension of unembryonated T. canis eggs. A suspension of T. canis eggs in 2% water agar was used as control group. The assay was done in triplicate for each fungus and the control group. SEM observations were performed on the 4th, 7th, 14th, 21st, and 28th day after inoculation. The effect of the fungi on eggs was evaluated in accordance with the alterations observed on the surface and the changes in the normal characteristics of the eggs. Hyphae around the eggs, appresoria penetrating the shell and changes in the typical egg membrane were observed in this assay. Type 3 effect (alterations that occur both in the embryo and the shell, and hyphal penetration of the eggs) was the prevalent effect. SEM allowed us to observe clearly the morphological alterations in T. canis eggs due to the effect of C. indicum and C. keratinophylum. Both saprophytic species of Chrysosporium alter the egg structure and alterations increase as exposure increases.

[Blastocystis spp.: Advances, controversies and future challenges]. / Blastocystis spp.: avances, controversias y desafíos futuros.

Blastocystis spp. is the most common protozoan detected in human stool samples. In developing countries, infection rates are higher than 20%. The presence of this parasite in the feces of several host species suggests its zoonotic potential. The clinical relevance and the pathogenic role of Blastocystis spp. in the intestinal tract remain unclear. There are several clinical reports that recognize it as the etiologic agent of several intestinal disorders such as diarrhea, inflammatory bowel disease and ulcerative colitis, although the pathogenicity of this parasite has not been proved yet. This wide range of clinical manifestations could be related to the genetic diversity exhibited by this parasite.

Bibliometric analysis of scientific literature on intestinal parasites in Argentina during the period 1985-2014.

The study of scientific production is a good indicator of the progress in research and knowledge generation. Bibliometrics is a scientific discipline that uses a set of indicators to quantitatively express the bibliographic characteristics of scientific publications. The scientific literature on the epidemiology of intestinal parasites in Argentina is scattered in numerous sources, hindering access and visibility to the scientific community. Our purpose was to perform a quantitative, bibliometric study of the scientific literature on intestinal parasites in humans in Argentina published in the period 1985-2014. This bibliometric analysis showed an increase in the number of articles on intestinal parasites in humans in Argentina published over the past 30 years. Those articles showed a collaboration index similar to that of the literature, with a high index of institutionality for national institutions and a very low one for international collaboration. The original articles were published in scientific journals in the American Continent, Europe and Asia. The use of bibliometric indicators can provide a solid tool for the diagnosis and survey of the research on epidemiology of intestinal parasites and contributes to the dissemination and visibility of information on the scientific production developed in Argentina.

In vitro and in vivo effects of Enterococcus faecalis CECT7121 on Toxocara canis.

The aim of the present paper was to evaluate the larvicidal effect of Enterococcus faecalis CECT7121 (Ef7121) on the Toxocara canis cycle both in vitro and in vivo. For the in vitro experiments, T. canis larvae were incubated with the supernatants of Ef7121 (EI) and mutant Ef7121 (EIm), in a pre-culture of Ef7121 (EII) and in a fresh culture with Ef7121 (EIII) and the Ef7121 mutant strain (EIIIm). The viability of the larvae was calculated after a 48 h incubation. A significant reduction of the viability of T. canis larvae was observed in EI, EII and EIII. A decrease of this inhibitory effect was observed in EIm and EIIIm (p = 0.008). In the in vivo experiments, mice were orally inoculated with three doses of Ef7121. To study the probiotic persistence in the intestine, the animals were sacrificed every four days and their intestines were dissected. The initial average bacterial levels were 9.7 x 10(4) for Ef7121 (colony forming units/g). At the end of the assay the levels were 1.46 x 10(4). No bacterial translocation was detected in mesenteric lymphatic nodules and spleen. Ef7121 interference with the biological cycle was evaluated in mice challenged with T. canis. The interference was significant when the mice were challenged with probiotic and T. canis simultaneously (p = 0.001), but it was not significant when the challenge was performed 15 days after administration of the bacterial inoculum (p = 0.06). In conclusion, Ef7121 possessed in vitro and in vivo larvicidal activity.

Detection of ß-Lactamase-Producing Enterococcus faecalis and Vancomycin-Resistant Enterococcus faecium Isolates in Human Invasive Infections in the Public Hospital of Tandil, Argentina.

The study's aim was to analyze the population structure of enterococci causing human invasive infections in a medium-sized Argentinian Hospital coincidental with a 5 year-period of increased recovery of antibiotic resistant enterococci (2010-2014). Species identification (biochemical testing/MALDI-TOF-MS), antimicrobial susceptibility (disk-diffusion) and clonal relatedness (PFGE/MLST/BAPS) were determined according to standard guidelines. ß-lactamase production was determined by a nitrocefin test and confirmed by PCR/sequencing. The isolates were identified as Enterococcus faecalis and Enterococcus faecium at a 2:1 ratio. Most of the E. faecalis isolates, grouped in 25 PFGE-types (ST9/ST179/ST236/ST281/ST388/ST604/ST720), were resistant to high-levels (HLR) of gentamicin/streptomycin. A ST9 clone (bla+/HLR-gentamicin) was detected in patients of different wards during 2014. E. faecium isolates were grouped in 10 PFGE-types (ST25/ST18/ST19/ST52/ST792), with a low rate of ampicillin resistance. Five vancomycin-resistant E. faecium, three vanA (ST792/ST25) and two vanB (ST25) were detected. The ST25 clone carried either vanA or vanB. The recovery of a bla+-ST9-E. faecalis clone similar to that described in the late 1980s in Argentina suggests the possibility of a local hidden reservoir. These results reflect the relevance of local epidemiology in understanding the population structure of enterococci as well as the emergence and spread of antimicrobial resistance in predominant enterococcal clonal lineages.

Cryptosporidium infection in calves from a rural area of Buenos Aires, Argentina.

Dairy calves less than 1 month of age are commonly infected with Cryptosporidium spp. The objective of the present study was to evaluate the prevalence of Cryptosporidium spp. among dairy calves or=810 oocysts/field. This study shows that Cryptosporidium spp. is one of the causes of calf neonatal diarrhoea in a rural area of Buenos Aires, Argentina. The highest intensity of infection reported for the

Heterogeneous distribution of human cystic echinococcosis after a long-term control program in Neuquén, Patagonia Argentina.

The present study is the first comprehensive analysis of human cystic echinococcosis (CE) epidemiological data carried out in the province of Neuquén, Patagonia Argentina, after 34 years of uninterrupted control program. A retrospective study of all the official records of confirmed human CE cases between 1995 and 2004 was carried out. A total of 1107 cases were reported. The overall mean annual incidence (MAI) was 24.4 per 100,000 inhabitants for the total population and 9.7/100,000 in the 0-14 year group. Distribution of cases by age and sex as well as frequency of cyst locations was analysed. Children accounted for 13.3% of total cases, indicating active transmission of the disease. Territorial distribution of cases was highly heterogeneous: MAI per 100,000 ranged from 7.9 in the Metropolitan Sanitary Area to 78.4 in the western rural areas. Higher values were recorded in small communities as Pilo Lil (800/100,000). MAI showed a significant decrease from 1995 to 1999 (43.9-18.8) but remains stable from 2000 to 2004 (15.9-15.5). These results suggest that standard control measures, despite long-term implementation, are not able to produce a sustained improvement of the epidemiological status of the disease. Further studies about local transmission cycles, definite and intermediate hosts present, Echinococcus granulosus strains or cultural behaviours in small communities are required in order to adequate the control actions in Neuquén.

Intestinal parasitoses and environmental factors in a rural population of Argentina, 2002-2003.

We evaluate the prevalence of intestinal parasites in 504 people and the degree of association between environmental variables and parasites found in population, soil and water in a rural area of Argentina during 2002-2003. A structured survey was used to evaluate the environmental variables and fecal-human, soil and water samples were analyzed. The prevalence of parasites was 45.4%. Most prevalent protozoa were Blastocystis hominis (27.2%) and Giardia lamblia (6.9%), while the most prevalent helminth was Ascaris lumbricoides (3.8%). The analyzed environmental variables showing association (p < 0.05) with presence of parasites in population were: cardboard-tin or wooden house, dirt floor, home or communal water pump, faucet outside the house or public faucet and cesspool or latrine. Parasite forms were found in 82.3% of the soil samples and in 84.2% of the water samples. In both samples we found parasites that were also found in people. In this study we have found deficient sanitary conditions associated with presence of parasites in population and we have evidenced that contaminated soil and water were the source of these parasites.

Reduction in surgical antibiotic prophylaxis expenditure and the rate of surgical site infection by means of a protocol that controls the use of prophylaxis.

OBJECTIVE: To evaluate the effectiveness of an intervention based on training and the use of a protocol with an automatic stop of antimicrobial prophylaxis to improve hospital compliance with surgical antibiotic prophylaxis guidelines. DESIGN: An interventional study with a before-after trial was conducted in 3 stages: a 3-year initial stage (January 1999 to December 2001), during which a descriptive-prospective survey was performed to evaluate surgical antimicrobial prophylaxis and surgical site infections; a 6-month second stage (January to June 2002), during which an educational intervention was performed regarding the routine use of a surgical antimicrobial prophylaxis request form that included an automatic stop of prophylaxis (the "automatic-stop prophylaxis form"); and a 3-year final stage (July 2002 to June 2005), during which a descriptive-prospective survey of surgical antimicrobial prophylaxis and surgical site infections was again performed. SETTING: An 88-bed teaching hospital in Entre Ríos, Argentina. PATIENTS: A total of 3,496 patients who underwent surgery were included in the first stage of the study and 3,982 were included in the final stage. RESULTS: Comparison of the first stage of the study with the final stage revealed that antimicrobial prophylaxis was given at the appropriate time to 55% and 88% of patients, respectively (relative risk [RR], 0.27 [95% confidence interval {CI}, 0.25-0.30]; P<.01); the antimicrobial regimen was adequate in 74% and 87% of patients, respectively (RR, 0.50 [95% CI, 0.45-0.55]; P<.01); duration of the prophylaxis was adequate in 44% and 55% of patients, respectively (RR, 0.80 [95% CI, 0.77-0.84]; P<.01); and the surgical site infection rates were 3.2% and 1.9%, respectively (RR, 0.59 [95% CI, 0.44-0.79]; P<.01). Antimicrobial expenditure was 10,678.66 US$ per 1,000 patient-days during the first stage and 7,686.05 US$ per 1,000 patient-days during the final stage (RR, 0.87 [95% CI, 0.86-0.89]; P<.01). CONCLUSION: The intervention based on training and application of a protocol with an automatic stop of prophylaxis favored compliance with the hospital's current surgical antibiotic prophylaxis guidelines before the intervention, achieving significant reductions of surgical site infection rates and substantial savings for the healthcare system.

Viability and infectiousness of eggs of Echinococcus granulosus aged under natural conditions of inferior arid climate.

We studied the viability and infectiousness of aged Echinococcus granulosus eggs by in vivo evaluation in ovines. Our results demonstrated that after 41 months of ageing of the eggs under environmental conditions of an inferior arid climate (Patagonia, Argentina), they were still able to produce infection in 4/4 ovines challenged with 1200 eggs per ovine. In the ovines experimentally infected with these aged eggs, the occurrence of hepatic and pulmonary cysts was determined by necropsy, histologic and genetic studies. The eggs were found in a semi-senescent stage, thus keeping their capacity to generate an infection in the intermediary ovine host.

Enterobius vermicularis infection among population of General Mansilla, Argentina.

AIM: To evaluate the relationships between the personal, sociocultural, and environmental characteristics, and the presence or absence of symptoms with the detection of Enterobius vermicularis (E. vermicularis) in a population sample in our region (General Mansilla, Province of Buenos Aires, Argentina), by individual and familiar analyses. METHODS: E. vermicularis was diagnosed in 309 people from 70 family units residing in the urban area and the rural area of the city of General Mansilla. Each of them was surveyed so as to register personal, environmental and sociocultural data. Questions about the presence or absence of anal itch, abdominal pain and sleeping disorder were also asked. Significant associations were determined by square chi tests. Logistic regression models were adjusted by using a backward conditional stepwise method to determine the presence of this parasite in the individuals and in the families. RESULTS: The parasites were found in 29.12% (90/309) of the individuals, with a frequency of 14.28% (20/140) among the heads of the families and of 41.42% (70/169) among the children. The only variables showing a significant association were affiliation, where the risk category was "being the son/daughter of", and the symptoms were abdominal pain, sleeping disorder, and anal itch. Families with a member infected with parasite were considered Positive Families (PF) and they were 40/70 (57.14%), only 5% (2/40) of the PF had 100% of their members infected with the parasite. The logistic regression models applied showed that the risk categories were mainly affiliation (son/daughter) and housing (satisfactory) among others. CONCLUSION: The presence of E. vermicularis was proved in one third of the studied population. The frequency of families with all their members infected with the parasite was very low. Most of the studied personal, sociocultural, and environmental variables did not turn out to be significantly associated with the presence of the parasite. An association with the category of "son/daughter" and housing classified as "satisfactory" was determined. The latter may be due to the fact that the people living in that category of housing have hygienic practices at home that favour the distribution of the eggs in the environment. The presence of the analysed symptoms was associated with the presence of the parasite, thus strengthening the need of periodical control of the population showing at least one of these symptoms.

Cryptosporidium parvum GP60 subtypes in dairy cattle from Buenos Aires, Argentina.

Cryptosporidium parvum from 73 dairy calves less than two months old from Buenos Aires province (Argentina) were molecularly characterized using sequence analysis of the GP60 gene. Seventy-five sequences were obtained, and seven different subtypes were identified, all belonging to the IIa subtype family. The most common subtypes were IIaA20G1R1 (27/75), IIaA22G1R1 (16/75), and IIaA18G1R1 (13/75). Subtypes IIaA21G1R1, IIaA23G1R1, IIaA16G1R1 and IIaA19G1R1 were found sporadically. Two samples contained mixed infections with IIaA21G1R1 and IIaA22G1R1. A significant association was found between subtypes and geographic location, whereas there was no relation between subtypes and presence of diarrhea. Three of the subtypes found in this study (IIaA16G1R1, IIaA18G1R1, and IIaA19G1R1) were previously identified in humans. These findings suggest that cattle could play an important role in the transmission of cryptosporidiosis to humans in Buenos Aires province.

Blastocystis spp.: avances, controversias y desafíos futuros / Blastocystis spp.: Advances, controversies and future challenges

Blastocystis spp. es el protista intracelular que se detecta con mayor frecuencia en muestras de materia fecal humana; las tasas de infección pueden superar el 20% en países en vías de desarrollo. El hallazgo de este parásito en heces de diversas especies animales sugiere su potencial zoonótico. La relevancia clínica y el papel patógeno de Blastocystis spp. en el tracto intestinal son inciertos. Varias son las publicaciones que lo reconocen como agente etiológico de desórdenes intestinales como diarrea, enfermedad inflamatoria intestinal y colitis ulcerosa, aunque la patogenicidad de este parásito no ha sido probada. Este amplio rango de respuestas a la infección podría estar relacionado con la diversidad genética de los aislamientos provenientes de hospedadores infectados.

Blastocystis spp. is the most common protozoan detected in human stool samples. In developing countries, infection rates are higher than 20%. The presence of this parasite in the feces of several host species suggests its zoonotic potential. The clinical relevance and the pathogenic role of Blastocystis spp. in the intestinal tract remain unclear. There are several clinical reports that recognize it as the etiologic agent of several intestinal disorders such as diarrhea, inflammatory bowel disease and ulcerative colitis, although the pathogenicity of this parasite has not been proved yet. This wide range of clinical manifestations could be related to the genetic diversity exhibited by this parasite.

Experimental infection with Cryptosporidium parvum IIaA21G1R1 subtype in immunosuppressed mice.

Cryptosporidium parvum subtype IIaA21G1R1 oocysts were used to infect dexamethasone immunosuppressed N: NIH Swiss mice. This is the first Cryptosporidium mouse model in which the relationship between infection and apoptosis has been histologically studied at each portion of the gut in order to observe this dynamic in chronic cryptosporidiosis. Histology showed developmental stages in the duodenum, proximal and distal jejunum, ileum, cecum and colon, with the small intestine remaining infected until day 35 post infection. At proximal jejunum an inverse correlation between infection and apoptosis was observed at days 28 and 35 p.i. Data suggests that jejunum could be an interesting place to carry out further studies on the dynamics of Cryptosporidium infection and apoptosis. Based on these findings, this mouse model was useful to evaluate clinical, parasitological and histological aspects of C. parvum subtype IIaA21G1R1 infection, and it will be an appropriate tool to investigate different aspects of Cryptosporidium infection.

[Community participation in the control of intestinal parasitoses at a rural site in Argentina]. / Participación comunitaria en el control de las parasitosis intestinales en una localidad rural de Argentina.

OBJECTIVES: To design, implement, and evaluate a complex plan of actions aimed at reducing parasitoses in a rural town in the province of Buenos Aires, Argentina, with the involvement of the community. METHODS: The project took place in General Mansilla, Buenos Aires province, in three stages. First, the initial epidemiological situation of intestinal parasitoses in the community was assessed by analyzing for parasites a series of stool samples and anal swabs taken from a sample of 522 individuals. Second, interventions were implemented: the pharmacological treatment of the infected individuals, and health education for the public involving local promoters. Third, the pharmacological treatment was evaluated with followup stool testing for those treated; the health education effort was evaluated through a survey and stool testing for parasites of those who had participated solely in the education. RESULTS: The frequency of intestinal parasitoses was 58.2%; of the total, 43.9% were protozoa and 35.2%, helminths. The most common pathogens were Enterobius vermicularis, Blastocystis hominis, and Giardia lamblia. The deworming treatment reduced intestinal parasitoses to 15.1% (P < 0.001), and was more effective in helminths than in protozoa. Intestinal parasitoses declined following the health education intervention, in general (from 58.2% to 47.9%; P = 0.019), as well as for helminths (from 35.2% to 20.3%; P < 0.001), and hygiene habits improved significantly. CONCLUSIONS: Treatment of parasitosis and the educational intervention provided by local promoters made it possible to reduce parasitic diseases-particularly those caused by helminths-in the community under study. We recommend extending this experience to other rural communities and broadening the interventions to cutoff additional modes of transmission, such as water and food.

First genotyping of Giardia lamblia from human and animal feces in Argentina, South America.

The purpose of this study was to investigate the genotypes of Giardia lamblia from human and animal feces and their epidemiological and clinical characteristics in Argentina, South America. Seventy isolates, 60 from humans (adults and children), eight from dogs and two from cows were processed by polymerase chain reaction-restriction fragment length polymorphism. Data corresponding to demographic, socio-cultural and environmental variables and presence/absence of signs/symptoms were collected. The triosephosphate isomerase gene was amplified from 43 (71.66%) of the 60 human fecal samples. Among these, 3/43 (6.98%) were genotype AII and 40/43 (93.02%) were genotype B. Assemblage AII was detected in three children who lived together in a shantytown and they were oligosymptomatic and none had diarrhea. This genotype was not found in animals. Genotype B showed a high prevalence in both adults and children. It was also found in polysymptomatic people, many of whom presented diarrhea. It was also found only in one dog. The present study represents the first contribution to the knowledge of G. lamblia genotypes in Argentina.

In vitro and in vivo effects of Enterococcus faecalis CECT7121 on Toxocara canis

The aim of the present paper was to evaluate the larvicidal effect of Enterococcus faecalis CECT7121 (Ef7121) on the Toxocara canis cycle both in vitro and in vivo. For the in vitro experiments, T. canis larvae were incubated with the supernatants of Ef7121 (EI) and mutant Ef7121 (EIm), in a pre-culture of Ef7121 (EII) and in a fresh culture with Ef7121 (EIII) and the Ef7121 mutant strain (EIIIm). The viability of the larvae was calculated after a 48 h incubation. A significant reduction of the viability of T. canis larvae was observed in EI, EII and EIII. A decrease of this inhibitory effect was observed in EIm and EIIIm (p = 0.008). In the in vivo experiments, mice were orally inoculated with three doses of Ef7121. To study the probiotic persistence in the intestine, the animals were sacrificed every four days and their intestines were dissected. The initial average bacterial levels were 9.7 x 10(4) for Ef7121 (colony forming units/g). At the end of the assay the levels were 1.46 x 10(4). No bacterial translocation was detected in mesenteric lymphatic nodules and spleen. Ef7121 interference with the biological cycle was evaluated in mice challenged with T. canis. The interference was significant when the mice were challenged with probiotic and T. canis simultaneously (p = 0.001), but it was not significant when the challenge was performed 15 days after administration of the bacterial inoculum (p = 0.06). In conclusion, Ef7121 possessed in vitro and in vivo larvicidal activity.

First genotyping of Giardia lamblia from human and animal feces in Argentina, South America

The purpose of this study was to investigate the genotypes of Giardia lamblia from human and animal feces and their epidemiological and clinical characteristics in Argentina, South America. Seventy isolates, 60 from humans (adults and children), eight from dogs and two from cows were processed by polymerase chain reaction-restriction fragment length polymorphism. Data corresponding to demographic, socio-cultural and environmental variables and presence/absence of signs/symptoms were collected. The triosephosphate isomerase gene was amplified from 43 (71.66 percent) of the 60 human fecal samples. Among these, 3/43 (6.98 percent) were genotype AII and 40/43 (93.02 percent) were genotype B. Assemblage AII was detected in three children who lived together in a shantytown and they were oligosymptomatic and none had diarrhea. This genotype was not found in animals. Genotype B showed a high prevalence in both adults and children. It was also found in polysymptomatic people, many of whom presented diarrhea. It was also found only in one dog. The present study represents the first contribution to the knowledge of G. lamblia genotypes in Argentina.

Intestinal parasitoses and environmental factors in a rural population of Argentina, 2002-2003

We evaluate the prevalence of intestinal parasites in 504 people and the degree of association between environmental variables and parasites found in population, soil and water in a rural area of Argentina during 2002-2003. A structured survey was used to evaluate the environmental variables and fecal-human, soil and water samples were analyzed. The prevalence of parasites was 45.4 percent. Most prevalent protozoa were Blastocystis hominis (27.2 percent) and Giardia lamblia (6.9 percent), while the most prevalent helminth was Ascaris lumbricoides (3.8 percent). The analyzed environmental variables showing association (p < 0.05) with presence of parasites in population were: cardboard-tin or wooden house, dirt floor, home or communal water pump, faucet outside the house or public faucet and cesspool or latrine. Parasite forms were found in 82.3 percent of the soil samples and in 84.2 percent of the water samples. In both samples we found parasites that were also found in people. In this study we have found deficient sanitary conditions associated with presence of parasites in population and we have evidenced that contaminated soil and water were the source of these parasites.

Hemos evaluado la prevalencia de parásitos intestinales en 504 personas y el grado de asociación entre las variables ambientales y los parásitos intestinales hallados en la población, el suelo y el agua de una zona rural de Argentina durante 2002-2003. Se utilizó una encuesta estructurada para relevar las variables ambientales y se examinaron muestras fecales humanas, de agua y de suelo. La prevalencia parasitaria fue de 45,4 por ciento. Los protozoos prevalentes fueron Blastocystis hominis (27,2 por ciento) y Giardia lamblia (6,9 por ciento), mientras que el helminto mas prevalente fue Ascaris lumbricoides (3,8 por ciento). Las variables ambientales analizadas que mostraron asociación (p < 0,05) con la presencia de parásitos en la población fueron: casa de cartón-chapa o de madera con piso de tierra, bomba de agua domiciliaria o comunitaria, canilla fuera de la casa o pública y pozo ciego o letrina. Elementos parasitarios fueron hallados en el 82,3 por ciento de las muestras de suelo y en el 84,2 por ciento de las muestras de agua. En ambas muestras fueron hallados parásitos que también se encontraron en las personas. En este trabajo hemos hallado deficientes condiciones sanitarias asociadas con la presencia de parásitos en las personas y hemos evidenciado que el suelo y el agua contaminada fueron la fuente de esos parasitos.

Comparación de métodos de lisis y extracción de ADN de trofozoítos de Giardia lamblia / Comparison of lysis methods and DNA extraction of Giardia lamblia trophozoites

Se evaluó la eficiencia de procedimientos de lisis y tratamientos de extracción de ADN de trofozoítos de Giardia lamblia respecto a la eficiencia de ruptura, cantidad y pureza de ADN, además de los tiempos de procesamiento y costos. Se testearon cinco métodos de lisis (agua destilada y calor; agua destilada, calor y proteinasa K; buffer de lisis D; buffer de lisis E y un kit comercial) y tres métodos de purificación de ADN (fenol:cloroformo: isoamílico; Chelex 100 y un kit comercial). Los datos obtenidos se analizaron estadísticamente. La combinación de buffer de lisis E y Chelex fue un método simple y económico, que produjo alto rendimiento de ADN con baja pureza. Ella técnica comercial fue un método simple, más costoso que produjo bajas cantidades de ADN con un nivel de pureza apropiado para estudios moleculares.