Genome-wide support for incipient Tula hantavirus species within a single rodent host lineage.

Evolutionary divergence of viruses is most commonly driven by co-divergence with their hosts or through isolation of transmission after host shifts. It remains mostly unknown, however, whether divergent phylogenetic clades within named virus species represent functionally equivalent byproducts of high evolutionary rates or rather incipient virus species. Here, we test these alternatives with genomic data from two widespread phylogenetic clades in Tula orthohantavirus (TULV) within a single evolutionary lineage of their natural rodent host, the common vole Microtus arvalis. We examined voles from forty-two locations in the contact region between clades for TULV infection by reverse transcription (RT)-PCR. Sequencing yielded twenty-three TULV Central North and twenty-one TULV Central South genomes, which differed by 14.9-18.5 per cent at the nucleotide and 2.2-3.7 per cent at the amino acid (AA) level without evidence of recombination or reassortment between clades. Geographic cline analyses demonstrated an abrupt (<1 km wide) transition between the parapatric TULV clades in continuous landscape. This transition was located within the Central mitochondrial lineage of M. arvalis, and genomic single nucleotide polymorphisms showed gradual mixing of host populations across it. Genomic differentiation of hosts was much weaker across the TULV Central North to South transition than across the nearby hybrid zone between two evolutionary lineages in the host. We suggest that these parapatric TULV clades represent functionally distinct, incipient species, which are likely differently affected by genetic polymorphisms in the host. This highlights the potential of natural viral contact zones as systems for investigating the genetic and evolutionary factors enabling or restricting the transmission of RNA viruses.

Exploring the influence of density-dependence and weather on the spatial and temporal variation in common vole (Microtus arvalis) abundance in Castilla y León, NW Spain.

BACKGROUND: The common vole has invaded the agroecosystems of northwestern Spain, where outbreaks cause important crop damage and management costs. Little is yet known about the factors causing or modulating vole fluctuations. Here, we used 11 years of vole abundance monitoring data in 40 sites to study density-dependence and weather influence on vole dynamics. Our objective was to identify the population dynamics structure and determine whether there is direct or delayed density-dependence. An evaluation of climatic variables followed, to determine whether they influenced vole population peaks. RESULTS: First- and second-order outbreak dynamics were detected at 7 and 33 study sites, respectively, together with second-order variability in periodicity (2-3 to 4-5-year cycles). Vole population growth was explained by previous year abundance (mainly numbers in summer and spring) at 21 of the sites (52.5%), by weather variables at 11 sites (27.5%; precipitation or temperature in six and five sites, respectively), and by a combination of previous abundance and weather variables in eight sites (20%). CONCLUSIONS: We detected variability in vole spatiotemporal abundance dynamics, which differs in cyclicity and period. We also found regional variation in the relative importance of previous abundances and weather as factors modulating vole fluctuations. Most vole populations were cyclical, with variable periodicity across the region. Our study is a first step towards the development of predictive modeling, by disclosing relevant factors that might trigger vole outbreaks. It improves decision-making processes within integrated management dealing with mitigation of the agricultural impacts caused by voles. © 2023 Society of Chemical Industry.

The oral bioavailability of soil-borne risk elements for small terrestrial mammals: Microtus arvalis (Pallas, 1778) and Apodemus sylvaticus L. and its implication in environmental studies.

The aims of this study were (i) to specify real risk elemetnt (RE) uptake by wild terresrial mammals (A. sylvaticus and M. arvalis), (ii) to describe RE distribution in critical organs such as the liver and kidney, and (iii) to determine potencial differences in RE toxicokinetics with regards to individual species or sex. Three groups of experimental animals were fed diets amended with soil and green biomass of hyperaccumulator Arabidopsis halleri with different RE levels. The contents of As, Cd, and Pb in the liver and kidneys of the animals reflected the element contents in the diet. Higher Cd and Pb accumulation ability was observed in A. sylvaticus compared to M. arvalis tissues, and an opposite pattern was recorded for As. Zn contents in tissues of both species remained unchanged, and total contents of Zn in the exposed animals even tended to decrease compared to the controls. Results of this study indicate a generally similar response of both species to elevated RE contents in the diet, confirming these wild rodents as suitable models for RE biomonitoring in ecosystems. However, our data highlights some distinction in As toxicokinetics in wood mice compared to that of field voles and a significantly higher accumulation of Pb and Cd in females. Therefore, factors of species and sex should not be overlooked if relevant data are to be obtained in environmental studies conducted on small terrestrial rodents.

Seasonal Adaptation: Geographic Photoperiod-Temperature Patterns Explain Genetic Variation in the Common Vole Tsh Receptor.

The vertebrate photoperiodic neuroendocrine system uses the photoperiod as a proxy to time the annual rhythms in reproduction. The thyrotropin receptor (TSHR) is a key protein in the mammalian seasonal reproduction pathway. Its abundance and function can tune sensitivity to the photoperiod. To investigate seasonal adaptation in mammals, the hinge region and the first part of the transmembrane domain of the Tshr gene were sequenced for 278 common vole (Microtus arvalis) specimens from 15 localities in Western Europe and 28 localities in Eastern Europe. Forty-nine single nucleotide polymorphisms (SNPs; twenty-two intronic and twenty-seven exonic) were found, with a weak or lack of correlation with pairwise geographical distance, latitude, longitude, and altitude. By applying a temperature threshold to the local photoperiod-temperature ellipsoid, we obtained a predicted critical photoperiod (pCPP) as a proxy for the spring onset of local primary food production (grass). The obtained pCPP explains the distribution of the genetic variation in Tshr in Western Europe through highly significant correlations with five intronic and seven exonic SNPs. The relationship between pCPP and SNPs was lacking in Eastern Europe. Thus, Tshr, which plays a pivotal role in the sensitivity of the mammalian photoperiodic neuroendocrine system, was targeted by natural selection in Western European vole populations, resulting in the optimized timing of seasonal reproduction.

Spatial and Temporal Dynamics and Molecular Evolution of Tula orthohantavirus in German Vole Populations.

Tula orthohantavirus (TULV) is a rodent-borne hantavirus with broad geographical distribution in Europe. Its major reservoir is the common vole (Microtus arvalis), but TULV has also been detected in closely related vole species. Given the large distributional range and high amplitude population dynamics of common voles, this host-pathogen complex presents an ideal system to study the complex mechanisms of pathogen transmission in a wild rodent reservoir. We investigated the dynamics of TULV prevalence and the subsequent potential effects on the molecular evolution of TULV in common voles of the Central evolutionary lineage. Rodents were trapped for three years in four regions of Germany and samples were analyzed for the presence of TULV-reactive antibodies and TULV RNA with subsequent sequence determination. The results show that individual (sex) and population-level factors (abundance) of hosts were significant predictors of local TULV dynamics. At the large geographic scale, different phylogenetic TULV clades and an overall isolation-by-distance pattern in virus sequences were detected, while at the small scale (<4 km) this depended on the study area. In combination with an overall delayed density dependence, our results highlight that frequent, localized bottleneck events for the common vole and TULV do occur and can be offset by local recolonization dynamics.

FREQUENT LEPTOSPIRA SPP. DETECTION BUT ABSENCE OF TULA ORTHOHANTAVIRUS IN MICROTUS SPP. VOLES, NORTHWESTERN SPAIN.

The common vole (Microtus arvalis) is a major agricultural pest in Europe and is a reservoir for several zoonotic agents, such as Leptospira spp. and Tula orthohantavirus (TULV). However, little is known about the occurrence of those pathogens in voles from Spain, where the species has largely expanded its distribution range in the past decades, causing agricultural pests and zoonotic diseases. For a molecular survey, 580 common voles and six Lusitanian pine voles (Microtus lusitanicus) were collected in 26 localities from four provinces of northwestern Spain. We assessed the presence of Leptospira spp. DNA in kidney tissue by PCR targeting the lipL32 gene, detecting a prevalence of 7.9% (95% confidence interval, 5.9-10.4) for common voles and of 33.3% (95% confidence interval, 4.3-77.7) for Lusitanian pine voles. We identified Leptospira kirschneri in 24 animals and Leptospira borgpetersenii in two animals, using secY gene-specific PCR. We analyzed environmental and demographic factors (such as age class, weight, and sex) and population dynamics data for their potential effect on the Leptospira spp. prevalence in those voles. The Leptospira spp. DNA detection rate in common voles increased significantly with maximum air temperature, vole weight, and amount of accumulated rainfall during the 90 d before capture and within the peak phase of the population cycle. We assessed the presence of TULV in lung tissue of 389 voles by reverse-transcription PCR, with no positive results. The absence of TULV might be explained by the evolutionary isolation of the common vole in Spain. The detection of two Leptospira genomospecies underlines the necessity for further typing efforts to understand the epidemiology of leptospiral infection in the common vole and the potential risk for human health in Spain.

Behavioural plasticity of motor personality traits in the common vole under three-day continual observation in a test box.

In animals, behavioural personality traits have been well-documented in a wide array of species. However, these traits, different between individuals, are not completely stable in individuals. They show behavioural plasticity like many other phenotypic traits. This plasticity is able to overcome some weak aspects of personality trait behavioural strategy. In the present study, we examined the relationship between motor personality traits and behavioural plasticity in the common vole (Microtus arvalis) using a PhenoTyper (PT) box (Noldus). During a three-day test, four behavioural motor activity parameters were monitored in 47 voles: distance moved, (loco)motion duration, motion change frequency, sprint duration. Consistency repeatability (RC) of the parameters from the PT test was very high, with all values ≥ 0.91. To select the best linear mixed-effect models (LMMs), several predictors (test day, sex, body weight) were tested. Only test day had a significant effect on the dependent variables and other predictors did not improve the LMMs. Further, we found significant effects of random intercepts (motor personality traits) and slopes (behavioural plasticity), as well as significant negative correlations between them for all behavioural parameters. Our results indicate that motor personality traits were connected with behavioural plasticity. Moreover, we revealed a significant positive correlation between the random slopes of (loco)motion duration and motion change frequency. This relationship could indicate some central plasticity of motor personality traits. In conclusion, negative correlations between the motor personality traits and the behavioural plasticity demonstrate expression of convergent tendency from both opposite trait values. This corresponds with different ideas on ability to compensate personality effects or to prepare for potential future conditions. In the laboratory, plasticity of personality traits take place whenever an animal is placed e. g. in a breeding box for the first time or is left for a long time in an experimental apparatus.

Hantavirus-Leptospira coinfections in small mammals from central Germany.

European orthohantaviruses (Puumala orthohantavirus (PUUV); Dobrava-Belgrade orthohantavirus (DOBV), genotype Kurkino; Tula orthohantavirus (TULV)), and Leptospira spp. are small mammal-associated zoonotic pathogens that cause diseases with potentially similar symptoms in humans. We investigated the frequency of Leptospira spp. and hantavirus single and double infections in small mammals from 22 sites in Thuringia, central Germany, during 2017. TULV infections were detected at 18 of 22 sites (mean prevalence 13.8%, 93/674). PUUV infections were detected at four of 22 sites (mean prevalence 1.5%, 7/471), and respective PUUV sequences formed a novel phylogenetic clade, but DOBV infections were not detected at all. Leptospira infections were detected at 21 of 22 sites with the highest overall prevalence in field voles (Microtus agrestis) with 54.5% (6/11) and common voles (Microtus arvalis) with 30.3% (205/676). Leptospira-hantavirus coinfections were found in 6.6% (44/671) of common voles but only in two of 395 bank voles. TULV and Leptospira coinfection probability in common voles was driven by individual (age) and population-level factors. Coinfections seemed to be particularly associated with sites where Leptospira spp. prevalence exceeded 35%. Future investigations should evaluate public health consequences of this strong spatial clustering of coinfections.

In Vivo Characterization of a Bank Vole-Derived Cowpox Virus Isolate in Natural Hosts and the Rat Model.

Cowpox virus (CPXV) belongs to the genus Orthopoxvirus in the Poxviridae family and is endemic in western Eurasia. Based on seroprevalence studies in different voles from continental Europe and UK, voles are suspected to be the major reservoir host. Recently, a CPXV was isolated from a bank vole (Myodes glareolus) in Germany that showed a high genetic similarity to another isolate originating from a Cotton-top tamarin (Saguinus oedipus). Here we characterize this first bank vole-derived CPXV isolate in comparison to the related tamarin-derived isolate. Both isolates grouped genetically within the provisionally called CPXV-like 3 clade. Previous phylogenetic analysis indicated that CPXV is polyphyletic and CPXV-like 3 clade represents probably a different species if categorized by the rules used for other orthopoxviruses. Experimental infection studies with bank voles, common voles (Microtusarvalis) and Wistar rats showed very clear differences. The bank vole isolate was avirulent in both common voles and Wistar rats with seroconversion seen only in the rats. In contrast, inoculated bank voles exhibited viral shedding and seroconversion for both tested CPXV isolates. In addition, bank voles infected with the tamarin-derived isolate experienced a marked weight loss. Our findings allow for the conclusion that CPXV isolates might differ in their replication capacity in different vole species and rats depending on their original host. Moreover, the results indicate host-specific differences concerning CPXV-specific virulence. Further experiments are needed to identify individual virulence and host factors involved in the susceptibility and outcome of CPXV-infections in the different reservoir hosts.

Patchy Occurrence of Cowpox Virus in Voles from Germany.

Cowpox virus (CPXV), genus Orthopoxvirus, family Poxviridae, is a zoonotic pathogen in Eurasian wild rodents. High seroprevalences have been reported previously for vole and murine species in Europe. In contrast, viral DNA was only rarely detected, and very few reservoir-derived CPXV isolates exist. In this study, CPXV DNA and CPXV-reactive antibodies were monitored in wild small mammals for 5 years in four German federal states. Screening of liver tissues of 3966 animals by CPXV real-time PCR (qPCR) revealed five voles of two species positive for CPXV DNA. Two positive bank voles (Myodes glareolus) and two positive common voles (Microtus arvalis) originated from two plots in Baden-Wuerttemberg. One positive bank vole originated from Mecklenburg-Western Pomerania. None of the small mammals from Thuringia and North Rhine-Westphalia was positive in the qPCR. CPXV antigen-based indirect immunofluorescence assays of 654 highly diluted chest cavity fluid samples detected two bank voles and two common voles from the same sites in Baden-Wuerttemberg to be highly seroreactive. Five animals were CPXV DNA positive, and four other animals were orthopoxvirus seropositive. Our study indicates both a very low prevalence and a patchy occurrence of CPXV in common and bank voles and absence in other rodent and shrew species in Germany. The multiple detection of infected voles at one site in Baden-Wuerttemberg and continued detection in a region of Mecklenburg-Western Pomerania classify these regions as potential endemic foci.

Concentration of Mercury in the Livers of Small Terrestrial Rodents from Rural Areas in Poland.

Small terrestrial mammals could be used as accumulative biomonitors of different environmental contaminants, but the knowledge of the level of Hg in their bodies is scant. The aim of our research was to verify the factors influencing Hg bioaccumulation and to analyze the concentration of total mercury (Hg) in the livers of four species of wild terrestrial rodents from different rural areas of Poland: the yellow-necked mouse (Apodemus flavicollis), striped field mouse (Apodemus agrarius), common vole (Microtus arvalis), and bank vole (Myodes glareolus). The concentration of total Hg was analyzed in liver tissue by atomic absorption spectrometry using a direct mercury analyzer. The concentration of Hg found in the livers of rodents ranged from <1 to 36.4 µg/kg of wet weight, differed between study sites, species, and sexes, and was related to body weight. We addressed feeding habits as potential causes of differences in liver Hg concentration among species.

Rodent Host Abundance and Climate Variability as Predictors of Tickborne Disease Risk 1 Year in Advance.

Using long-term data on incidences of Lyme disease and tickborne encephalitis, we showed that the dynamics of both diseases in central Europe are predictable from rodent host densities and climate indices. Our approach offers a simple and effective tool to predict a tickborne disease risk 1 year in advance.

Numerical response of a mammalian specialist predator to multiple prey dynamics in Mediterranean farmlands.

The study of rodent population cycles has greatly contributed, both theoretically and empirically, to our understanding of the circumstances under which predator-prey interactions destabilize populations. According to the specialist predator hypothesis, reciprocal interactions between voles and small predators that specialize on voles, such as weasels, can cause multiannual cycles. A fundamental feature of classical weasel-vole models is a long time-lag in the numerical response of the predator to variations in prey abundance: weasel abundance increases with that of voles and peaks approximately 1 yr later. We investigated the numerical response of the common weasel (Mustela nivalis) to fluctuating abundances of common voles (Microtus arvalis) in recently colonized agrosteppes of Castilla-y-Léon, northwestern Spain, at the southern limit of the species' range. Populations of both weasels and voles exhibited multiannual cycles with a 3-yr period. Weasels responded quickly and numerically to changes in common-vole abundance, with a time lag between prey and weasel abundance that did not exceed 4 months and occurred during the breeding season, reflecting the quick conversion of prey into predator offspring and/or immigration to sites with high vole populations. We found no evidence of a sustained, high weasel abundance following vole abundance peaks. Weasel population growth rates showed spatial synchrony across study sites approximately 60 km apart. Weasel dynamics were more synchronized with that of common voles than with other prey species (mice or shrews). However, asynchrony within, as well as among sites, in the abundance of voles and alternative prey suggests that weasel mobility could allow them to avoid starvation during low-vole phases, precluding the emergence of prolonged time lag in the numerical response to voles. Our observations are inconsistent with the specialist predator hypothesis as currently formulated, and suggest that weasels might follow rather than cause the vole cycles in northwestern Spain. The reliance of a specialized predator on a functional group of prey such as small rodents does not necessarily lead to a long delay in the numerical response by the predator, depending on the spatial and interspecific synchrony in prey dynamics.

Detection of Francisella tularensis in three vole species in Central Europe.

Francisella tularensis is a zoonotic, gram-negative bacterium that causes tularemia in humans. Depending on its subspecies and the route of transmission, mild to lethal courses have been reported. F. tularensis subsp. holarctica is the only subspecies found in Europe and affects a plenitude of vertebrates including lagomorphs and rodents. Population outbreaks of certain rodent species are likely to be involved in the transmission of this pathogen. This molecular survey aims to evaluate the presence of F. tularensis in small mammals from three Central European countries. Using a real-time polymerase chain reaction, F. tularensis DNA was detected in common voles (Microtus arvalis) from Switzerland and in field voles (Microtus agrestis) and a bank vole (Myodes glareolus) from Germany, but not in any other small mammal species. All common voles from the Czech Republic were negative for F. tularensis DNA. The prevalence in the three vole species varied between 1.3% and 3.0%. In conclusion, Francisella tularensis DNA was detected in three vole species in two of three countries investigated. The observed low prevalence raises questions on the role of voles for the transmission of Francisella tularensis in Central Europe.

Influence of no-tillage versus tillage system on common vole (Microtus arvalis) population density.

BACKGROUND: While the 'no-tillage' management system generally improves soil properties and helps to control arthropod pests, it may also intensify crop infestation by the common vole (Microtus arvalis Pallas). In this study, we evaluated the impact of soil management (no-tillage or tillage), crop and previous crop (winter wheat or winter rape), and season (spring or autumn) on common vole density using data from the Common Vole Monitoring Programme undertaken by the Plant Protection Service of the Czech Republic between 2000 and 2009. RESULTS: Models predicted low mean values of vole infestation across management types, crops, and seasons. The untilled fields hosted significantly more voles than the tilled fields in spring but not in autumn. More common voles were found in winter rape than in winter wheat during both seasons. CONCLUSION: Recent studies suggest that no-tillage management is more profitable than tillage management as a result of its positive impact on soil properties and pest control. During periods of high vole infestation, however, tillage may constitute an alternative strategy for reducing yield losses. © 2017 Society of Chemical Industry.

The complete mitochondrial genome of the common vole, Microtus arvalis (Rodentia: Arvicolinae).

The common vole, Microtus arvalis belongs to the genus Microtus in the subfamily Arvicolinae. In this study, the complete mitochondrial genome of M. arvalis was recovered using shotgun sequencing and an iterative mapping approach using three related species. Phylogenetic analyses using the sequence of 21 arvicoline species place the common vole as a sister species to the East European vole (Microtus levis), but as opposed to previous results we find no support for the recognition of the genus Neodon within the subfamily Arvicolinae, as this is, as well as the genus Lasiopodomys, found within the Microtus genus.

Epidemiological Investigations of Four Cowpox Virus Outbreaks in Alpaca Herds, Germany.

Four cowpox virus (CPXV) outbreaks occurred in unrelated alpaca herds in Eastern Germany during 2012-2017. All incidents were initially noticed due to severe, generalized, and finally lethal CPXV infections, which were confirmed by testing of tissue and serum samples. As CPXV-infection has been described in South American camelids (SACs) only three times, all four herds were investigated to gain a deeper understanding of CPXV epidemiology in alpacas. The different herds were investigated twice, and various samples (serum, swab samples, and crusts of suspicious pox lesions, feces) were taken to identify additionally infected animals. Serum was used to detect CPXV-specific antibodies by performing an indirect immunofluorescence assay (iIFA); swab samples, crusts, and feces were used for detection of CPXV-specific DNA in a real-time PCR. In total, 28 out of 107 animals could be identified as affected by CPXV, by iIFA and/or PCR. Herd seroprevalence ranged from 16.1% to 81.2%. To investigate the potential source of infection, wild small mammals were trapped around all alpaca herds. In two herds, CPXV-specific antibodies were found in the local rodent population. In the third herd, CPXV could be isolated from a common vole (Microtus arvalis) found drowned in a water bucket used to water the alpacas. Full genome sequencing and comparison with the genome of a CPXV from an alpaca from the same herd reveal 99.997% identity, providing further evidence that the common vole is a reservoir host and infection source of CPXV. Only in the remaining fourth herd, none of the trapped rodents were found to be CPXV-infected. Rodents, as ubiquitous reservoir hosts, in combination with increasingly popular alpacas, as susceptible species, suggest an enhanced risk of future zoonotic infections.

Isolation and complete genome characterization of novel reassortant orthoreovirus from common vole (Microtus arvalis).

A novel mammalian orthoreovirus (MRV) strain was isolated from the lung tissue of a common vole (Microtus arvalis) with Tula hantavirus infection. Seven segments (L1-L3, M2-M3, S2, and S4) of the Hungarian MRV isolate MORV/47Ma/06 revealed a high similarity with an MRV strain detected in bank vole (Myodes glareolus) in Germany. The M1 and S3 segment of the Hungarian isolate showed the closest relationship with the sequence of a Slovenian human and a French murine isolate, respectively. The highest nucleotide and amino acid identity values were above 90 and 95% in all of the comparisons to the reference sequences in GenBank, except for the S1 with a maximum of 69.6% nucleotide and 75.4% amino acid identity. As wild rodents are among the main sources of zoonotic infections, the reservoir role of these animals and zoonotic potential of rodent origin MRVs need to be further investigated.

Evidence for selection maintaining MHC diversity in a rodent species despite strong density fluctuations.

Strong spatiotemporal variation in population size often leads to reduced genetic diversity limiting the adaptive potential of individual populations. Key genes of adaptive variation are encoded by the immune genes of the major histocompatibility complex (MHC) playing an essential role in parasite resistance. How MHC variation persists in rodent populations that regularly experience population bottlenecks remains an important topic in evolutionary genetics. We analysed the consequences of strong population fluctuations on MHC class II DRB exon 2 diversity in two distant common vole (Microtus arvalis) populations in three consecutive years using a high-throughput sequencing approach. In 143 individuals, we detected 25 nucleotide alleles translating into 14 unique amino acid MHC alleles belonging to at least three loci. Thus, the overall allelic diversity and amino acid distance among the remaining MHC alleles, used as a surrogate for the range of pathogenic antigens that can be presented to T-cells, are still remarkably high. Both study populations did not show significant population differentiation between years, but significant differences were found between sites. We concluded that selection processes seem to be strong enough to maintain moderate levels of MHC diversity in our study populations outcompeting genetic drift, as the same MHC alleles were conserved between years. Differences in allele frequencies between populations might be the outcome of different local parasite pressures and/or genetic drift. Further understanding of how pathogens vary across space and time will be crucial to further elucidate the mechanisms maintaining MHC diversity in cyclic populations.

Rodent food quality and its relation to crops and other environmental and population parameters in an agricultural landscape.

The diet, its quality and quantity considerably influence population parameters of rodents. In this study, we used NIRS methods for estimation of nitrogen content in stomachs of rodent populations. The study was carried out in diverse arable landscape in South Moravia, Czech Republic. Rodents were sampled in cultural crops (alfalfa, barley, wheat, sunflower, maize and rape) as well as in fallow habitats (herbal set-aside and old orchard). Influence of habitat, date, year, individual parameters (body length, sex, breeding and age), and relative abundance on quality of consumed food was studied. Under conditions of higher population density, dominant species [wood mouse (Apodemus sylvaticus) and common vole (Microtus arvalis)] consumed food richer in nitrogen. Also the strong effect of crop and date (season) was found in both species. There was no significant effect of the other parameters studied on food quality (N-content).