A simple assay for measuring tannin-protein precipitation capacity offers insights into the diet and food choice of black howler monkeys (Alouatta pigra).

Phenolics, like tannins, are plant-specialized metabolites that play a protective role against herbivory. Tannins can reduce palatability and bind with proteins to reduce digestibility, acting as deterrents to feeding and impacting nutrient extraction by herbivores. Some assays measure tannin and total phenolics content in plants but lack determination of their biological effects, hindering the interpretation of tannin function in herbivory and its impacts on animal behavior and ecology. In this study, we successfully applied the radial diffusion assay to assess tannin protein precipitation (PP) capacity and evaluate the anti-nutritional effects of tannins in food plants (n = 24) consumed by free-ranging black howler monkeys (Alouatta pigra) in Tabasco, Mexico. We found PP rings in five plant species consumed by the monkeys. The mature fruit of Inga edulis was the most consumed food plant, despite having a high tannin PP capacity (56.66 mg tannic acid equivalent/g dry matter). These findings highlight the presence of tannins in the black howler diet and provide insight into the primates' resilience and potential strategies for coping with anti-nutritional aspects of the diet.

Host traits, ownership behaviour and risk factors of SARS-CoV-2 infection in domestic pets in Mexico.

SARS-CoV-2 can infect pets under natural conditions, which raises questions about the risk factors related to the susceptibility of these animals to infection. The status of pet infection by SARS-CoV-2 in Mexico is not well-understood. We aimed to estimate the frequency of positive household cats and dogs to viral RNA and antibodies for SARS-CoV-2 during the second wave of human infections in Mexico, and to recognize the major risk factors related to host and pet ownership behaviour. We evaluated two study groups, cats and dogs from COVID-19-infected/-suspected households (n = 44) and those admitted for veterinary care for any reason at several veterinary hospitals in Puebla City, Mexico (n = 91). Using RT-PCR, we identified the presence of SARS-CoV-2 RNA in swabs of four dogs (18.18%) and zero cats in COVID-19-infected/-suspected households; within this group, 31.82% of dogs and 27.27% of cats were tested IgG ELISA-positive; and neutralizing antibodies were detected in one dog (4.55%) and two cats (9.09%). In the random group (pets evaluated at private clinics and veterinary teaching hospital), 25.00% of dogs and 43.59% of cats were ELISA-positive and only one cat showed neutralizing antibodies (2.56%). Older than 4-year-old, other pets at home, and daily cleaning of pet dish, were each associated with an increase in SARS-CoV-2 infection (p < 0.05). Allowing face lick, sharing bed/food with pets and owner tested positive or suspected COVID-19 were not significant risk factors, but more than 4 h the owner spent away from home during the lockdown for COVID-19 (OR = 0.37, p = 0.01), and outdoor pet food tray (OR = 0.32, p = 0.01) significantly decreased the risks of SARS-CoV-2 infection in pets, suggesting that time the owner spends with their pet is an important risk factor.

Influence of tannic acid concentration on the physicochemical characteristics of saliva of spider monkeys (Ateles geoffroyi).

Tannins are a chemical defense mechanism of plants consumed by herbivores. Variations in salivary physicochemical characteristics such as pH, total protein concentration (TP), and presence of proline-rich proteins (PRPs) in animals have been reported as a mechanism to protect the oral cavity when consuming food with variations in pH and tannins. Variations in salivary physiochemistry as adaptations for consuming tannin-rich foods have been found in omnivorous and folivorous primates, but have not yet been reported in frugivorous species such as spider monkeys. We therefore assessed changes in pH using test strips, TP concentration by measuring absorbance at 595 nm in a spectrophotometer and salivary PRPs using the SDS-PAGE electrophoresis technique in the saliva of nine captive spider monkeys in response to the consumption of solutions with different concentrations of tannic acid. The results showed variations in pH, TP concentration and the presence and variation of possible salivary PRPs associated with tannic acid concentration. These findings suggest that spider monkeys may tailor their salivary physicochemical characteristics in response to the ingestion of potentially toxic compounds.

Salivary proteome of a Neotropical primate: potential roles in host defense and oral food perception.

BACKGROUND: Saliva contains a very complex mixture of proteins for defense against microbiological pathogens and for oral food perception. Howler monkeys are Neotropical primates that can consume a mostly leaf diet. They are well known to thrive in highly disturbed habitats where they may cope with a diversity of dietary challenges and infection risks. We aimed to describe the salivary proteome of howlers to contribute to better understanding of their physiology. METHODS: We analyzed the salivary proteins of wild black howler monkeys (Alouatta pigra), by SDS-PAGE-1-D and Nano LC-MS/MS and categorized them by their function involved in host defense and oral food perception. RESULTS: Our proteomic analysis identified 156 proteins in howler saliva including a number of host defense peptides that are the first line of defense in mammals, such as defensin, cathelicidin, dermcidin, and lactotransferrin, and proteins with anti-bacterial, anti-fungal, and anti-viral capacity, such as IgA, IgG, IgM, BPI, salivary heat shock 70 kDa protein, beta-2-microbulin, and protein S-100. We also identified key proteins necessary for taste perception, including salivary carbonic anhydrase VI, cystatin D, IgA, and fatty acid-binding protein. Proteins to detect astringent foods were identifying, including four members of cystatins (A, B, C and D), lactoperoxidase, and histidine-rich proteins. No chitinase and amylase were identified as would be expected because howlers do not eat insects and little starch. These findings provide basic information to future studies in oral biology, ingestive physiology, and physiological ecology of mammals and non-human primates.

Salivary tannin-binding proteins are a pervasive strategy used by the folivorous/frugivorous black howler monkey.

Dietary tannins can affect protein digestion and absorption, be toxic, and influence food selection by being astringent and bitter tasting. Animals that usually ingest tannins may regularly secrete tannin-binding salivary proteins (TBSPs) to counteract the negative effects of tannins or TBSPs production can be induced by a tannin-rich diet. In the wild, many primates regularly eat a diet that contains tannin-rich leaves and unripe fruit and it has been speculated that they have the physiological ability to cope with dietary tannins; however, details of their strategy remains unclear. Our research details the salivary protein composition of wild and zoo-living black howler monkeys (Alouatta pigra) feeding on natural versus manufactured low-tannin diets, and examines differences in TBSPs, mainly proline-rich proteins (PRPs), to determine whether production of these proteins is dependent on the tannin content of their food. We measured the pH, flow rate, and concentration of total protein and trichloroacetic acid soluble proteins (an index of PRPs) in saliva. Howler monkeys produced slightly alkaline saliva that may aid in the binding interaction between tannin and salivary proteins. We used gel electrophoresis to describe the salivary protein profile and this analysis along with a tannin-binding assay allowed us to detect several TBSPs in all individuals. We found no differences in the characteristics of saliva between wild and zoo-living monkeys. Our results suggest that black howler monkeys always secrete TBSPs even when fed on foods low in tannins. This strategy of constantly using this salivary anti-tannin defense enables them to obtain nutrients from plants that sometimes contain high levels of tannins and may help immediately to overcome the astringent sensation of their food allowing howler monkeys to eat tanniferous plants.