Anemia in Children Aged Four to Eight from a Semirural Community in Central East Area of Argentina.

We present the results of the first stage of the Program for the Control of Intestinal Parasitosis and Nutrition, analyzing the frequency of anemia and its relation with intestinal parasitic infections and socio-cultural and environmental factors present in school children from a semirural community of Argentina. A total of 123 children aged 4-8 years were interviewed; 93 (75.6%) of them showed up for blood extraction and a fecal sample was taken properly. The frequency of anemia was 33.3%; 45.0% in children aged 4-5 years and 24.5% in those aged 6-8 years (p=0.038). Intestinal parasites were found in 83.9% of the children; 75.0% in children aged 4-5 years and 90.6% in those aged 6-8 years (p=0.043). No statistical differences were found when relating parasitic infections, social/cultural variables and housing characteristic with anemia, probably of nutritional origin. This study reveals the coexistence of anemia and parasitic infections in apparently healthy children who were unnoticed by the conventional public health system.

Nutritional status in parasitized and nonparasitized children from two districts of Buenos Aires, Argentina.

UNLABELLED: The Program for the Control of Intestinal Parasites and Nutrition was designed to intervene in small communities to prevent and control the effects of parasitic infections on children's health. OBJECTIVES: To analyze the association between nutritional status and parasitic infection in suburban and rural children from Buenos Aires, Argentina. METHODS: Nutritional status was assessed by anthropometric (weight, height, BMI, skinfolds, upper arm circumference, muscle, and fat upper arm areas) and biochemical (Hb, Ca, Mg, Zn, and Cu) indicators. Parasitological analysis were made on both serial stool and perianal swab samples. A total of 708 children aged 3-11 were measured. The biochemical analysis included 217 blood samples and the parasitological study included 284 samples. RESULTS: Anthropometric status was similar in both settings with low rates of underweight and stunting (<6%), and high rates of overweight (~17%) and obesity (~12%). Ca deficiency was significantly higher in suburban children where 80% of them were hypocalcemic. Around 70% of fecal samples contained parasites. Among infected children, the most prevalent species were Blastocystis hominis and Enterobius vermicularis (~43%) followed by Giardia lamblia (~17%). Differences in parasitological status between districts were not significant. In the suburban district parasitized children were lighter, shorter, and had a lower upper arm circumference than their non-infected peers. No differences in anthropometric status were seen among infected and uninfected rural children. CONCLUSIONS: The results suggest an association between intestinal parasites and physical growth in suburban children. Rural children seem to be protected against the effects of parasitic infection.

Intestinal parasitoses in suburban and rural school children in Argentina

Neste trabalho, são apresentados os resultados da primeira etapa do Programa de controle de doençasparasitárias intestinais e nutrição (PROCOPIN) e comparadas as condições de saúde pública e os hábitos associados a parasitos intestinais em crianças de zona suburbana (SC) e de zona rural (SR) na Argentina. Recolheu-se informação pessoal e sobre as condições de saúde pública. Foram realizados exames parasitológicos de fezes e de esfregaço perianal com gazes seriados. Foram examinadas 465 SC e 251 RC, entre as quais se encontraram os seguintes índices de infecção: em SC, 39,1por cento e em RC, 31,1por cento, sendo p igual 0,032. Os parasitos mais frequentes em ambas as localizações foram: Blastocystis hominis, Enterobius vermicularis e Giardia intestinalis. Más condições de saúde pública nas casas de SC não constituíram fator de risco de a criança ser parasitada, o que sugereque a transmissão desses parasitos não ocorre na casa, mas, provavelmente, em escolas, clubes ou outras áreas de recreação que congregam grande número de crianças. Entre as RC, condições de saúde pública dentro ou no entorno da casa apareceram como fator de risco, indicando que é principalmente ali onde ocorre a transmissão do parasito. Portanto, o comportamento social das crianças, as condições da saúde pública e os hábitos de higiene das populações devem ser vistoscomo fatores de risco para a parasitose.

Intestinal parasites and genotypes of Giardia intestinalis in school children from Berisso, Argentina.

INTRODUCTION: Intestinal parasitic infections have been reported in different regions of Argentina. Giardia intestinalis is recognized as "the national parasite". The aim of this work was to determine the prevalence of both intestinal parasites and G. intestinalis genotypes, as well as to analyze the clinical and epidemiological characteristics in schoolchildren from a suburban community. METHODOLOGY: Serial coproparasitological analysis and perianal swab method were performed in 244 schoolchildren. Demographic, sociocultural and environmental variables were registered. The presence of signs/symptoms and risk behaviours were also recorded. Stools with G. intestinalis were selected for genotyping. RESULTS: Out of 244 schoolchildren, 179/244 (73.4%) were infected with intestinal parasites. The presence of intestinal parasitosis was associated only with house flooding. Multivariate analysis identified that use of a latrine is significantly correlated with G. intestinalis and age six to 11 years with E. vermicularis. Signs and symptoms were recorded in 62% of infected children and in 57.9% of those not infected. Genomic amplification was revealed that 65.7% (46/70) of Giardia positive samples corresponded to genotype B, 31.4% (22/70) to genotype AII, and two samples (2.8%) had mixed infection (AII + B). CONCLUSIONS: This study shows a high percentage of infected children living in a suburban community in poor sanitary conditions, and not visiting the doctor in spite of evident signs and symptoms associated a digestive pathology. This situation supports the need for continuing the development of community programs allowing the improvement of quality of life and control of parasitosis in deprived populations.

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.

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.

Influencing factors in the occurrence of injured coliforms in the drinking water distribution system in the city of La Plata, Argentina.

The objective of this study was to evaluate the influencing factors in the occurrence of coliforms in the drinking water in La Plata (Argentina) from June 1999 to June 2001. A total of 180 samples were collected from Rio de La Plata (102 samples) and Puelche Aquifer (78 samples); 45 samples were collected for each of the four seasons. The membrane filter procedure was used for isolating bacteria, and each sample was tested for chlorine and pH. The highest percentage of samples positive for coliforms in the two tested media was obtained in summer while the highest percentage of negative samples was obtained in winter. No Escherichia coli was isolated. The percentage of injured coliforms fluctuated between 70 and 100%. The most frequently isolated bacteria was Enterobacter cloacae in summer, Enterobacter agglomerans in autumn and Klebsiella oxytoca in winter and spring. Significant correlations were observed between coliforms and the distance from the initial treatment point, and with the level of free chlorine. We conclude that drinking water contamination in La Plata occurs in the distribution system due to increased temperatures and reduced disinfectant levels, which result in bacterial regrowth.

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

OBJETIVOS: Diseñar, implementar y evaluar un plan complejo de acciones dirigido a reducir las parasitosis en una localidad rural de la provincia de Buenos Aires, Argentina, con la participación de la comunidad. MÉTODOS: El trabajo se realizó en General Mansilla, provincia de Buenos Aires, Argentina, en tres etapas. 1) Se evaluó la situación epidemiológica inicial de las parasitosis intestinales en la comunidad mediante análisis coproparasitológico seriado y de escobillado anal en una muestra de 522 personas. 2) Se implementaron dos intervenciones: el tratamiento farmacológico de las personas parasitadas y la educación sanitaria de la población con la participación activa de divulgadores locales. 3) El tratamiento antiparasitario se evaluó mediante análisis coproparasitológico de seguimiento de todas las personas tratadas; la intervención educativa se evaluó mediante una encuesta y análisis coproparasitológico a personas que solo habían participado en el plan de educación sanitaria. RESULTADOS: La frecuencia de parasitosis intestinal fue de 58,2 por ciento; del total, 43,9 por ciento por protozoos y 35,2 por ciento por helmintos. Los patógenos más frecuentes fueron Enterobius vermicularis, Blastocystis hominis y Giardia lamblia. El tratamiento antiparasitario redujo la parasitosis intestinal a 15,1 por ciento (P < 0,001), más eficaz en helmintos que en protozoos. La parasitosis intestinal disminuyó después de la intervención educativa sanitaria, tanto en sentido general (de 58,2 por ciento a 47,9 por ciento; P = 0,019) como por helmintos (de 35,2 por ciento a 20,3 por ciento; P < 0,001) y se mejoraron significativamente los hábitos higiénicos. CONCLUSIONES: El tratamiento parasitológico y la intervención educacional mediante divulgadores locales permitieron reducir las parasitosis en la comunidad estudiada, especialmente las provocadas por helmintos. Se recomienda extender esta experiencia a otras comunidades rurales y ampliarla con intervenciones...

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 percent; of the total, 43.9 percent were protozoa and 35.2 percent, helminths. The most common pathogens were Enterobius vermicularis, Blastocystis hominis, and Giardia lamblia. The deworming treatment reduced intestinal parasitoses to 15.1 percent (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 percent to 47.9 percent; P = 0.019), as well as for helminths (from 35.2 percent to 20.3 percent; 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.

[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.

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%. 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.

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.

PCR amplification of triosephosphate isomerase gene of Giardia lamblia in formalin-fixed feces.

Giardia lamblia (syn G. intestinalis, G. duodenalis) is the intestinal protozoan producing non-bacterial diarrhea most prevalent in the world. PCR genotype classification of Giardia in feces depends on the quality and quantity of purified DNA and the removal of a great number of inhibitors. The aim of this study was to adapt a PCR protocol to the amplification of the triosephosphate isomerase (tpi) gene of Giardia lamblia in formalin-fixed feces. The tpi gene of G. lamblia was amplified in 28 of the 34 analyzed samples (82.35%) and the B genotype was obtained in all cases. Two major modifications were implemented to improve the performance of PCR from formolated fecal matter. One of these improvements was the use of polyvinylpyrrolidone (PVP) and the other was the addition of bovine serum albumin (BSA). The PCR protocol used in this study showed an amplification percentage exceeding the values reported by other authors with high sensibility and specificity.

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.

Related factors to human toxocariasis in a rural community of Argentina.

The objective of this study was to evaluate the relationship between toxocariasis frequency and demographic, environmental, sanitary variables, eosinophylia, and other intestinal parasites in a rural population of Argentina. Serological examination of 100 individuals was carried out by using ELISA technique for the detection of anti-toxocara antibodies. Eosinophiles in peripheral blood, presence of intestinal parasites, and demographic, environmental, and socio-cultural data were evaluated. Eighty-one feces samples of dogs belonging to the studied people were analyzed to detect eggs of Toxocara canis. Thirty of them were from 30 dogs and 51 were pools from dog feces. Samples of dirt from around the homes (n: 47) and from public park (n: 4) were taken. To determine the associations, the c(2) and Fisher tests were used. The seroprevalence was 23%. Eosinophilia in peripheral blood was detected in 86.95% seropositive individuals and in 37.66% seronegative individuals (p < 0.001, OR = 11.03). Of the 23 people with positive serology, 69.56% had at least one intestinal parasite. All individuals with positive serology had dogs in their homes. Among the dog owners there was a significant association between the presence of anti-toxocara antibodies and home flooding. Eggs of T. canis were detected in the feces of 5/81 dogs and three of these dogs belonged to individuals with positive serology. Eggs of Toxocara spp. were found in 41.17% of the dirt samples, eight of which came from the area surrounding the homes of individuals with positive serology (p = 0.032; OR = 4.36). Taking into account all the variables influencing the frequency of toxocariasis in this population, the implementation of Public Health programs specifically focused on anti-parasitic treatment of dogs is recommended.

Related factors to human toxocariasis in a rural community of Argentina

The objective of this study was to evaluate the relationship between toxocariasis frequency and demographic, environmental, sanitary variables, eosinophylia, and other intestinal parasites in a rural population of Argentina. Serological examination of 100 individuals was carried out by using ELISA technique for the detection of anti-toxocara antibodies. Eosinophiles in peripheral blood, presence of intestinal parasites, and demographic, environmental, and socio-cultural data were evaluated. Eighty-one feces samples of dogs belonging to the studied people were analyzed to detect eggs of Toxocara canis. Thirty of them were from 30 dogs and 51 were pools from dog feces. Samples of dirt from around the homes (n: 47) and from public park (n: 4) were taken. To determine the associations, the c² and Fisher tests were used. The seroprevalence was 23 percent. Eosinophilia in peripheral blood was detected in 86.95 percent seropositive individuals and in 37.66 percent seronegative individuals (p < 0.001, OR = 11.03). Of the 23 people with positive serology, 69.56 percent had at least one intestinal parasite. All individuals with positive serology had dogs in their homes. Among the dog owners there was a significant association between the presence of anti-toxocara antibodies and home flooding. Eggs of T. canis were detected in the feces of 5/81 dogs and three of these dogs belonged to individuals with positive serology. Eggs of Toxocara spp. were found in 41.17 percent of the dirt samples, eight of which came from the area surrounding the homes of individuals with positive serology (p = 0.032; OR = 4.36). Taking into account all the variables influencing the frequency of toxocariasis in this population, the implementation of Public Health programs specifically focused on anti-parasitic treatment of dogs is recommended.