Antimicrobial potential of the Mayan medicine plant Matayba oppositifolia (A. Rich.) Britton against antibiotic-resistant priority pathogens.

ETHNOPHARMACOLOGICAL RELEVANCE: The bark of Matayba oppositifolia (A. Rich.) Britton (commonly known as "huaya" or "palo huacax") is commonly utilized in traditional Mayan medicine for treating diarrhea and for canker and other sores. AIM OF THE STUDY: The aim of this study was to investigate the in-vitro antimicrobial activity of M. oppositifolia bark extracts against drug-susceptible and -resistant ESKAPE-E pathogens. In addition, the phytochemical composition of the best antibacterial extract was analyzed. MATERIALS AND METHODS: The bark extracts were prepared with different solvents, including water, n-hexane, ethyl acetate and methanol. These were tested against ESKAPE-E (Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa and Enterobacter spp., including Escherichia coli) strains using Resazurin Microtiter Assay. In addition, the composition of the most active extract was analyzed by GC-MS. RESULTS: The aqueous and organic bark extracts showed activity on drug-susceptible and -resistant ESKAPE-E microbes (MIC = 1000-31.25 µg/mL). The n-hexane bark extract was more active against the superbugs carbapenem-resistant K. pneumoniae (MIC = 500-31.25 µg/mL) and A. baumannii (MIC = 250-125 µg/mL). The GC-MS analysis of this extract allowed the identification of 12 phytochemicals as the potential antibacterial compounds. The major compounds identified were palmitic acid (1), friedelan-3-one (2) and 7-dehydrodiosgenin (3). CONCLUSION: The present study reveals the strong in-vitro antibacterial activity of the n-hexane extract from the bark of M. oppositifolia and demonstrates the potential of natural products as a source of antibacterial compounds or phytomedicines that are specifically effective against drug-resistant ESKAPE-E bugs. Additionally, our investigation contributes to the ethnopharmacological knowledge and reappraisal of Mayan medicinal flora, as well as supports the traditional use of the bark of the medicinal plant M. oppositifolia for the treatment of infectious diseases.

Antibacterial and antibiofilm activities of Mayan medicinal plants against Methicillin-susceptible and -resistant strains of Staphylococcus aureus.

ETHNOPHARMACOLOGICAL RELEVANCE: Several medicinal plants are used in Mayan Traditional Medicine to treat skin, urinary, respiratory, and gastrointestinal infectious diseases. However, scientific studies that have supported the bioactivity of these Mayan medicinal plants are limited. AIM OF THE STUDY: To assess the in-vitro anti-Staphylococcus aureus growth and biofilm-formation activities of 15 Mayan medicinal plants that were selected based on their traditional uses for the treatment of infectious diseases. MATERIALS AND METHODS: Mayan medicinal plants used traditionally to treat infectious diseases were preselected. For each part of the plants, four extracts were prepared with different solvents (water, n-hexane, ethyl acetate, and methanol). These were tested against two reference strains: a Methicillin-susceptible and -resistant S. aureus, and two clinical isolates, including a susceptible and multidrug-resistant S. aureus using a Resazurin Microtiter Assay. In addition, the plant extracts were evaluated in biofilm-formation inhibition on S. aureus by means of the Crystal Violet method. RESULTS: A total of 120 extracts from 15 Mayan medicinal plant species belonging to 12 different families were selected according their ethnopharmacological uses to treat infectious diseases. Among the selected plant species, 26 extracts obtained from eight medicinal Mayan plants exhibited significant anti-S. aureus against the four strains tested. The most active extracts were the Aq (aqueous) leaf extract of Krugiodendron ferreum (Minimal Inhibitory Concentration [MIC] = 125-250 µg/mL), the MeOH bark extracts of Matayba oppositifolia, Clusia flava, Gymnopodium floribundum, the MeOH leaf extract of Spondias purpurea with MIC values of 250 µg/mL, and the MeOH leaf and Aq bark extracts of K. ferreum (MIC = 250-500 µg/mL). Among the active extracts, 12 exhibited a bactericidal effect on S. aureus strains (Minimal Bactericidal Concentration [MBC] = 250-1000 µg/mL). Forty extracts from 13 plants have an effect on the anti-formation of biofilm, the most active were the MeOH leaf extract of M. oppositifolia (one-half Inhibitory Concentration [IC50] = 10.4 µg/mL) and the MeOH (IC50 = 17.7 µg/mL) and Hex (18.2 µg/mL) leaf extracts from S. purpurea. CONCLUSION: Aqueous and organic extracts from Mayan medicinal plants showed bactericidal and anti-biofilm activities even against drug-resistant S. aureus strains. The present study supports the traditional usage of some plants employed in Mayan medicine for illnesses such as skin, gastrointestinal, and urinary infections and suggest that these plants could be a good source of antibacterial phytochemicals.

Rickettsia parkeri (Rickettsiales: Rickettsiaceae) detected in Amblyomma maculatum ticks collected on dogs in Tabasco, Mexico.

The present study was carried out to identify Rickettsia species with zoonotic potential in ticks collected from dogs in a rural area in Tabasco, Mexico. In total 197 Amblyomma maculatum ticks were collected from 40 domestic dogs. The collected specimens were pooled and subjected to DNA extraction. A fragment (380 bp) of citrate synthase gene (gltA) was amplified by polymerase chain reaction (PCR) using universal primers for Rickettsia. A second PCR was later performed to amplify a fragment (420 bp) of the outer membrane protein B gene (ompB). The PCR products were purified, sequenced and compared using the basic local alignment search tool (BLAST). Twenty out of 40 (50%) tick pools assayed were positive for rickettsial DNA using both primer pairs. The consensus sequence obtained from the ompB gene fragments showed 99.5-100% of identity with strains of Rickettsia parkeri. This study provides the first molecular evidence of the presence of R. parkeri in A. maculatum ticks infesting domestic dogs from southeastern Mexico. Close contact between dogs and humans should lead to consider the infection caused by this species of Rickettsia among the differential diagnoses for people of Tabasco, Mexico, who show acute febrile syndrome associated to inoculation eschar and have a clinical history of tick exposure.

Presence of Rickettsia Species in a Marginalized Area of Yucatan, Mexico.

In the state of Yucatan, Mexico, rickettsiosis has become a common vector-borne disease in the general population. Ectoparasite species such as Rhipicephalus sanguineus and Amblyomma mixtum have been identified as Rickettsia vectors in Yucatan by studies focused on the wild animal population in rural areas. There have been studies that have tried to determine the presence of Rickettsia species in ectoparasites collected in Yucatan, but these studies did not include marginalized areas, where living in close contact with domestic and peridomestic animals that carry ectoparasites is a high-risk factor for acquiring rickettsial infection or many other vector-borne diseases. We evaluated the vector diversity and the presence of Rickettsia species presence in the ectoparasite population that parasitizes domestic animals in a marginalized rural town of Yucatan, Mexico; we also evaluated the seroprevalence of rickettsial antibodies in the human population of this town in order to determine the prevalence of rickettsial infection. A total of 437 ectoparasites were collected from the study area. The tick specimens collected belonged to the species Rhipicephalus sanguineus (n=380, 49 positive), Amblyomma mixtum (n=3, 0 positive), Ixodes affinis (n=4, 0 positive), Ctenocephalides felis (n=33, 0 positive), and Trichodectes canis (n=17, 0 positive). Conventional polymerase chain reaction and sequencing were used to identify the DNA of Rickettsia. Six out of 354 (1.8%) serum samples were positive for antibody to R. typhi. The combination of low antibody titers and the presence of Rickettsia species infecting ectoparasite species found in the study area requires eco-epidemiological studies and the identification of potentially protective practices or habits.

Temporal distribution and genetic variants in influenza A(H1N1)pdm09 virus circulating in Mexico, seasons 2012 and 2013.

The 2012 and 2013 annual influenza epidemics in Mexico were characterized by presenting different seasonal patterns. In 2012 the A(H1N1)pdm09 virus caused a high incidence of influenza infections after a two-year period of low circulation; whereas the 2013 epidemic presented circulation of the A(H1N1)pdm09 virus throughout the year. We have characterized the molecular composition of the Hemagglutinin (HA) and Neuraminidase (NA) genes of the A(H1N1)pdm09 virus from both epidemic seasons, emphasizing the genetic characteristics of viruses isolated from Yucatan in Southern Mexico. The molecular analysis of viruses from the 2012 revealed that all viruses from Mexico were predominantly grouped in clade 7. Strikingly, the molecular characterization of viruses from 2013 revealed that viruses circulating in Yucatan were genetically different to viruses from other regions of Mexico. In fact, we identified the occurrence of two genetic variants containing relevant mutations at both the HA and NA surface antigens. There was a difference on the temporal circulation of each genetic variant, viruses containing the mutations HA-A141T / NA-N341S were detected in May, June and July; whereas viruses containing the mutations HA-S162I / NA-L206S circulated in August and September. We discuss the significance of these novel genetic changes.

Serological survey of Leptospira interrogans, Toxoplasma gondii and Trypanosoma cruzi in free roaming domestic dogs and cats from a marginated rural area of Yucatan Mexico.

To evaluate the serological status for Trypanosoma cruzi, Toxoplasma gondii and Leptospira interrogans antibodies in free roaming dogs and cats from a marginated rural community in Yucatan Mexico, 100 households were visited and animals sampled. From the 106 samples, 93 were from dogs and 13 were from cats. Frequency of positive results for T. gondii, T. cruzi and Leptospira spp was 97.8%, 9.7% and 45.2% for dogs and 92.3%, 0.0% and 15.2% for cats, respectively. No associations with age, sex and body condition was found for T. gondii and Leptospira spp neither for the place where pets sleep, fumigation or presence of triatomes in the household in the case of T. cruzi. For leptospirosis the most common serovars found were Canicola, Autralis and Bratislava in dogs and cats with titres of 100 or 200 with exception of one dog with a titre of 400. The high frequency of seropositive dogs suggests a high circulation of the agents in the population of free roaming owned dogs and cats probably due to the lack of control of the reservoirs and vectors involved. Domestic animals in those rural communities can be sentinels to assess the risk of human exposure in the rural communities.

Toxoplasma gondii presence in women with spontaneous abortion in Yucatan, Mexico.

The objective of this study was to estimate the seroprevalence of anti- Toxoplasma gondii antibodies and DNA of women with spontaneous abortions in 2 hospitals located in Yucatan, Mexico. Between June 2008 and May 2009, blood samples were taken from 100 women with spontaneous abortion attending the Ticul City Communitarian Hospital and the Merida Mother-Kid Hospital. The sera were tested for anti- T. gondii IgG and IgM antibodies. Blood samples (5 ml with anticlotting agent) were also used for polymerase chain reaction (PCR) assay, to detect T. gondii DNA. Forty-two of the 100 samples were negative. Of the positive samples (n = 58), 32 were positive to IgG, 2 to IgM, 5 to IgG and IgM, 6 to IgG and PCR, 1 to IgM and PCR, and 12 to IgG, IgM, and PCR. Accordingly, 55% of the women were seropositive to at least IgG, 20% to at least IgM, and 19% via PCR. Differences between hospitals were significant (P < 0.05) only for IgM. The risk of infection (IgM positive) was 2.85 (odds ratio [OR] 95%, confidence interval [CI]; 1.03-7.87) times greater in women patients at the Merida Mother-Kid Hospital, than those at the Ticul Communitarian Hospital. More studies are needed to evaluate the impact of this disease and to establish strategies to follow in order to reduce congenital toxoplasmosis in the populations at risk.

Entamoeba histolytica: cDNAs cloned as 30kDa collagen-binding proteins (CBP) belong to an antioxidant molecule family. Protection of hamsters from amoebic liver abscess by immunization with recombinant CBP.

A cDNA expression library of Entamoeba histolytica was screened with antiserum to native amoebic collagen binding proteins (CBPs), and two clones C13 and C7 which partially encode for the 30 kDa CBP were obtained. The sequenced clones were 90% homologous. C7 had a 69 bp deletion at the 5' end that is present in C13 and encodes for a Glu-Cys-Lys rich region and a four amino acids repeat (Glu-Lys-Glu-Cys). Purified fusion proteins from these cDNA clones were able to bind native type I collagen gels in a pH, calcium, ionic strength, and temperature dependent way. The binding of pgtC13 to collagen gel was time and temperature stable, while pgtC7 binding was not, suggesting that the deleted region in C7 is important for the binding. The clones reported here partially encode a 30 kDa CBP that also belong to an antioxidant molecule family. We demonstrated that the fusion protein pgtC13 is immunogenic and partially protective as a subunit vaccine in the hamster model of amoebic liver abscess.

Clinical-epidemiological study of leptospirosis in humans and reservoirs in Yucatan, Mexico

A leptospirosis clinical-epidemiological study was made in humans and reservoirs in the state of Yucatßn, MÚxico. Interviews and serological analyses were made on 400 persons from an open population, 439 probable cases of leptospirosis and 1060 animal reservoirs (cows, pigs, dogs, rats and opossums). IgM Leptospira DipstickTM and Microscopic Agglutination Test (MAT) was used to detect human antibodies to leptospiras and serovar respectively. Leptospirosis incidence in humans was 2.2/100,000 inhab. in 1998, 0.7/100,000 in 1999 and 0.9/100,000 in 2000. Overall seroprevalence was 14.2 percent, relatively unchanged from seroprevalences observed 20 years ago. Highest seropositivity was found in people over 56 years of age, predominating males over females. Predominant serovars in the open population were tarassovi, hardjo, pomona and panama. Leptospirosis cases were most frequent in rural areas, and the anicteric course predominated over the icteric. The panama, icterohaemorrhagiae and pomona serovars predominated in both anicteric and icteric courses. Dogs, pigs and rodents had the highest seropositivity among the reservoirs. Contact with rodents and natural water sources were significant factors (p ú 0.05). Human cases (74 percent) occurred during the rainy season. It is concluded that leptospirosis is still a serious illness with important clinical and epidemiological implications in the state of Yucatßn, Mexico

Clinical-epidemiological study of leptospirosis in humans and reservoirs in Yucatán, México.

A leptospirosis clinical-epidemiological study was made in humans and reservoirs in the state of Yucatán, México. Interviews and serological analyses were made on 400 persons from an open population, 439 probable cases of leptospirosis and 1060 animal reservoirs (cows, pigs, dogs, rats and opossums). IgM Leptospira Dipstick and Microscopic Agglutination Test (MAT) was used to detect human antibodies to leptospiras and serovar respectively. Leptospirosis incidence in humans was 2.2/100,000 inhab. in 1998, 0.7/100,000 in 1999 and 0.9/100,000 in 2000. Overall seroprevalence was 14.2%, relatively unchanged from seroprevalences observed 20 years ago. Highest seropositivity was found in people over 56 years of age, predominating males over females. Predominant serovars in the open population were tarassovi, hardjo, pomona and panama. Leptospirosis cases were most frequent in rural areas, and the anicteric course predominated over the icteric. The panama, icterohaemorrhagiae and pomona serovars predominated in both anicteric and icteric courses. Dogs, pigs and rodents had the highest seropositivity among the reservoirs. Contact with rodents and natural water sources were significant factors (p < or = 0.05). Human cases (74%) occurred during the rainy season. It is concluded that leptospirosis is still a serious illness with important clinical and epidemiological implications in the state of Yucatán, Mexico.