Role of Guinea Pigs (Cavia porcellus) Raised as Livestock in Ecuadorian Andes as Reservoirs of Zoonotic Yeasts

Guinea pigs (Cavia porcellus) have been reared for centuries in the Andean region for ceremonial purposes or as the main ingredient of traditional foods. The animals are kept in close proximity of households and interact closely with humans; this also occurs in western countries, where guinea pigs are considered pets. Even though it is acknowledged that domestic animals carry pathogenic yeasts in their tissues and organs that can cause human diseases, almost nothing is known in the case of guinea pigs. In this work we used traditional microbiological approaches and molecular biology techniques to isolate, identify, and characterize potentially zoonotic yeasts colonizing the nasal duct of guinea pigs raised as livestock in Southern Ecuador (Cañar Province). Our results show that 44% of the 100 animals studied were colonized in their nasal mucosa by at least eleven yeast species, belonging to eight genera: Wickerhamomyces, Diutina, Meyerozyma, Candida, Pichia, Rhodotorula, Galactomyces, and Cryptococcus. Noticeably, several isolates were insensitive toward several antifungal drugs of therapeutic use, including fluconazole, voriconazole, itraconazole, and caspofungin. Together, our results emphasize the threat posed by these potentially zoonotic yeasts to the farmers, their families, the final consumers, and, in general, to public and animal health.

Climate change, melting cryosphere and frozen pathogens: Should we worry ?

Permanently frozen environments (glaciers, permafrost) are considered as natural reservoirs of huge amounts of microorganisms, mostly dormant, including human pathogens. Due to global warming, which increases the rate of ice-melting, approximately 4 × 1021 of these microorganisms are released annually from their frozen confinement and enter natural ecosystems, in close proximity to human settlements. Some years ago, the hypothesis was put forward that this massive release of potentially-pathogenic microbes-many of which disappeared from the face of the Earth thousands and even millions of years ago-could give rise to epidemics. The recent anthrax outbreaks that occurred in Siberia, and the presence of bacterial and viral pathogens in glaciers worldwide, seem to confirm this hypothesis. In that context, the present review summarizes the currently available scientific evidence that allows us to imagine a near future in which epidemic outbreaks, similar to the abovementioned, could occur as a consequence of the resurrection and release of microbes from glaciers and permafrost. Supplementary Information: The online version of this article (10.1007/s42398-021-00184-8) contains supplementary material, which is available to authorized users.

Perspectives for using glacial and periglacial microorganisms for plant growth promotion at low temperatures.

Even though they are among the most extreme environments in which life can develop, glaciers are colonized by metabolically active microbes, some of which thrive-in their own particular way-under the prevailing harsh conditions. Glacial or periglacial microbes are often psychrophiles since they are able to grow optimally at low temperatures. This ability has evolved through a series of adaptations, both molecular and physiological, some of which have been exploited by the biotechnological industry to develop useful products and processes. The recent discovery of cold-adapted plant growth-promoting microorganisms (PGPM) in glacial ice or periglacial soils has opened a gate to a new trove of applications due to their potential use as biofertilizers or biocontrol agents, effective in cold climates. It has been claimed that this would be of profit to increase agriculture productivity in hilly terrains, like those prevailing in the Andes or the Himalayas, since-in addition to their ability to promote plant growth through direct or indirect mechanisms-they represent an environmentally friendly alternative to the use of pesticides and chemical fertilizers. In the following chapter, I summarize the current knowledge on the identity and characteristics of such PGPM and highlight the experiences in promoting the growth of a few plant species, at low temperatures.Key Points•Countless microbes are immured in glaciers and their surroundings.•Many glacial and periglacial microbes are cold-loving (i.e., psychrophiles).•Some glacial and periglacial psychrophiles promote plant growth and development.•Plant growth-promoting psychrophiles can be used to develop biofertilizers.

Bioprospecting cold-adapted plant growth promoting microorganisms from mountain environments.

Mountain soils are challenging environments for all kinds of living things, including plants and microorganisms. Many cold-adapted microorganisms colonizing these extreme soils play important roles as promoters of plant growth and development; for that reason, they are called collectively plant growth-promoting microorganisms (PGPM). Even though there is seldom doubt concerning the usefulness of PGPM to develop eco-friendly bioinoculants, including biofertilizers and biocontrollers, a series of aspects need to be addressed in order to make this technology field-applicable. Among these aspects, the ecological and rhizosphere competences of PGPM are of paramount importance, particularly when considering the development of bioinoculants, well suited for the intensification of mountainous agricultural production. Studies on native, cold-adapted PGPM conducted in the Indian Himalayan region (IHR) and the Tropical Andes (TA) lead nowadays the research in this field. Noticeably, some common themes are emerging. For instance, soils in these mountain environments are colonized by many cold-adapted PGPM able to mobilize soil nutrients and to inhibit growth of plant pathogens. Studies aimed at deeply characterizing the abilities of such PGPM is likely to substantially contribute towards a better crop productivity in mountainous environments. The present review focuses on the importance of this microbial resource to improve crop productivity in IHR and TA. We also present a number of successful examples, which emphasize the effectiveness of some bioinoculants-developed from naturally occurring PGPM-when applied in the field.

Diversity of culturable bacteria recovered from Pico Bolívar's glacial and subglacial environments, at 4950 m, in Venezuelan tropical Andes.

Even though tropical glaciers are retreating rapidly and many will disappear in the next few years, their microbial diversity remains to be studied in depth. In this paper we report on the biodiversity of the culturable fraction of bacteria colonizing Pico Bolívar's glacier ice and subglacial meltwaters, at ∼4950 m in the Venezuelan Andean Mountains. Microbial cells of diverse morphologies and exhibiting uncompromised membranes were present at densities ranging from 1.5 × 104 to 4.7 × 104 cells/mL in glacier ice and from 4.1 × 105 to 9.6 × 105 cells/mL in subglacial meltwater. Of 89 pure isolates recovered from the samples, the majority were eurypsychrophilic or stenopsychrophilic, according to their temperature range of growth. Following analysis of their 16S rDNA nucleotidic sequence, 54 pure isolates were assigned to 23 phylotypes distributed within 4 different phyla or classes: Beta- and Gammaproteobacteria, Actinobacteria, and Bacteroidetes. Actinobacteria dominated the culturable fraction of glacier ice samples, whereas Proteobacteria were dominant in subglacial meltwater samples. Chloramphenicol and ampicillin resistance was exhibited by 73.07% and 65.38%, respectively, of the subglacial isolates, and nearly 35% of them were multiresistant. Considering the fast rate at which tropical glaciers are melting, this study confirms the urgent need to study the microbial communities immured in such environments.

Bioprospecting glacial ice for plant growth promoting bacteria.

Glaciers harbor a wide diversity of microorganisms, metabolically versatile, highly tolerant to multiple environmental stresses and potentially useful for biotechnological purposes. Among these, we hypothesized the presence of bacteria able to exhibit well-known plant growth promoting traits (PGP). These kinds of bacteria have been employed for the development of commercial biofertilizers; unfortunately, these biotechnological products have proven ineffective in colder climates, like the ones prevailing in mountainous ecosystems. In the present work, we prospected glacial ice collected from two small tropical glaciers, located above 4.900 m in the Venezuelan Andes, for cold-active PGP bacteria. The initial screening strategy allowed us to detect the best inorganic-P solubilizers at low temperatures, from a sub-sample of 50 bacterial isolates. Solubilization of tricalcium phosphate, aluminum- and iron-phosphate, occurred in liquid cultures at low temperatures and was dependent on medium acidification by gluconic acid production, when bacteria were supplied with an appropriate source of carbon. Besides, the isolates were psychrophilic and in some cases exhibited a broad range of growth-temperatures, from 4 °C to 30 °C. Additional PGP abilities, including phytohormone- and HCN production, siderophore excretion and inhibition of phytopathogens, were confirmed in vitro. Nucleotidic sequence analysis of 16S rRNA genes allowed us to place the isolates within the Pseudomonas genus. Our results support the possible use of these strains to develop cold-active biofertilizers to be used in mountainous agriculture.

Bacteria recovered from a high-altitude, tropical glacier in Venezuelan Andes.

Glacial-ice microorganisms are intensively studied world-wide for a number of reasons, including their psychrophilic lifestyle, their usefulness in biotechnology procedures and their relationship with the search of life outside our planet. However, because of the difficulties for accessing and working at altitudes of >5.000 m above sea level, tropical glaciers have received much less attention than their arctic and antarctic counterparts. In the present work we isolated and characterized a total of forty-five pure isolates originating from direct plating of melted ice collected at the base of a rapidly-retreating, small glacier located at around 4.900 m.a.s.l. in Mount Humboldt (Sierra Nevada National Park, Mérida State, Venezuela). Initial examination of melted ice showed the presence of abundant- (>106 cells ml⁻¹), morphologically diverse- and active bacterial cells, many of which were very small ("dwarf cells"). The majority of the isolates were psychrophilic or psychrotolerant and many produced and excreted cold-active extracellular enzymes (proteases and amylases). The antibiotic tests showed an elevated percentage of isolates resistant to high doses (100 µg/ml) of different antibiotics including ampicillin, penicillin, nalidixic acid, streptomycin, chloramphenicol, kanamycin and tetracycline. Multiresistance was also observed, with 22.22 % of the strains simultaneously resistant up to five of the antibiotics tested. Metal resistance against Ni⁺⁺, Zn⁺⁺ and Cu⁺⁺ was also detected. In accordance with these results, plasmids of low and high molecular weight were detected in 47 % of the isolates. Twenty-two partial 16S rDNA sequences analyzed allowed grouping the isolates within five different phyla/classes: Alpha-, Beta- and Gamma-proteobacteria, Actinobacteria and Flavobacteria. This is the first report concerning South American Andean glacial ice microorganisms.

Characterization of the mineral phosphate-solubilizing activity of Pantoea agglomerans MMB051 isolated from an iron-rich soil in southeastern Venezuela (Bolívar State).

The mineral phosphate-solubilizing (MPS) activity of a Pantoea agglomerans strain, namely MMB051, isolated from an iron-rich, acidic soil near Ciudad Piar (Bolívar State, Venezuela), was characterized on a chemically defined medium (NBRIP). Various insoluble inorganic phosphates, including tri-calcium phosphate [Ca(3)(PO(4))(2)], iron phosphate (FePO(4)), aluminum phosphate (AlPO(4)), and Rock Phosphate (RP) were tested as sole sources of P for bacterial growth. Solubilization of Ca(3)(PO(4))(2) was very efficient and depended on acidification of the external milieu when MMB051 cells were grown in the presence of glucose. This was also the case when RP was used as the sole P source. On the other hand, the solubilization efficiency toward more insoluble mineral phosphates (FePO(4) and AlPO(4)) was shown to be very low. Even though gluconic acid (GA) was detected on culture supernatants of strain MMB051, a consequence of the direct oxidation pathway of glucose, inorganic-P solubilization seemed also to be related to other processes dependent on active cell growth. Among these, proton release by ammonium (NH(4)(+) ) fixation appeared to be of paramount importance to explain inorganic-P solubilization mediated by strain MMB051. On the contrary, the presence of nitrate (NO(3)(-) ) salts as the sole N source affected negatively the ability of MMB051 cells to solubilize inorganic P.

Mercury resistance in bacterial strains isolated from tailing ponds in a gold mining area near El Callao (Bolívar State, Venezuela).

Bacterial resistance to mercury (Hg) was investigated in strains isolated from Hg-contaminated tailing ponds located in the gold mining area of El Callao (Bolívar State, Venezuela). High frequencies of resistance were detected to both inorganic-Hg and organomercurials among these strains. A broad range of resistance levels was observed when determining minimal inhibitory concentrations of Hg(2+). Some strains were able to grow in liquid medium containing 25 muM: Hg(2+), whereas others grew at 300 muM: Hg(2+). Of 190 Hg-resistant strains tested, 58.2% were additionally shown to be resistant to ampicillin (40 mg/L), 33.3% to chloramphenicol (30 mg/L), 24.9% to streptomycin (30 mg/L), 23.3% to tetracycline (30 mg/L), and 1.6% to kanamycin (30 mg/L). Furthermore, we found that 20% of the Hg-resistant strains were simultaneously resistant to as many as four of these antibiotics, at the concentrations tested. The presence of large plasmids in 62.9% of 53 Hg-resistant strains screened prompted us to investigate the horizontal transfer of resistance determinants. Mating experiments were performed using Escherichia coli and Pseudomonas aeruginosa as recipient strains. The results obtained confirmed that indigenous Hg-resistant bacteria colonizing the tailing ponds can effectively transfer the phenotype to potentially pathogenic species.