Wolf Canis Lupus Research Articles

First report of occurrence of Babesia gibsoni in captive Indian wolves

Blood samples from fifteen captive Indian wolves (Canis lupus pallipes) maintained at Arignar Anna Zoological Park, Vandalur, Chennai were screened for the presence of Babesia spp., Ehrlichia canis and Trypnosoma evansi DNA by PCR. Out of 15 wolf samples, 3 samples were found positive for Babesia spp. The amplified 18S rRNA gene fragments from 3 wolves were sequenced and confirmed as Babesia gibsoni. A maximum likelihood tree was constructed using the three sequences along with other Babesia spp. sequences derived from GenBank adopting HKY nucleotide substitution model based on the Bayesian Information Criterion. The phylogenetic analysis confirmed that the three sequences were of Babesia gibsoni and highly divergent from Babesia canis, B. vogeli and B. vulpes. This might be a possible spill over event of B. gibsoni from community dogs through blood feeding dog ticks. This is the first report and molecular confirmation of B. gibsoni infection in captive Indian wolves.

White-tailed deer population dynamics in a multipredator landscape shaped by humans.

Large terrestrial mammals increasingly rely on human-modified landscapes as anthropogenic footprints expand. Land management activities such as timber harvest, agriculture, and roads can influence prey population dynamics by altering forage resources and predation risk via changes in habitat, but these effects are not well understood in regions with diverse and changing predator guilds. In northeastern Washington state, USA, white-tailed deer (Odocoileus virginianus) are vulnerable to multiple carnivores, including recently returned gray wolves (Canis lupus), within a highly human-modified landscape. To understand the factors governing predator-prey dynamics in a human context, we radio-collared 280 white-tailed deer, 33 bobcats (Lynx rufus), 50 cougars (Puma concolor), 28 coyotes (C. latrans), and 14 wolves between 2016 and 2021. We first estimated deer vital rates and used a stage-structured matrix model to estimate their population growth rate. During the study, we observed a stable to declining deer population (lambda = 0.97, 95% confidence interval: 0.88, 1.05), with 74% of Monte Carlo simulations indicating population decrease and 26% of simulations indicating population increase. We then fit Cox proportional hazard models to evaluate how predator exposure, use of human-modified landscapes, and winter severity influenced deer survival and used these relationships to evaluate impacts on overall population growth. We found that the population growth rate was dually influenced by a negative direct effect of apex predators and a positive effect of timber harvest and agricultural areas. Cougars had a stronger effect on deer population dynamics than wolves, and mesopredators had little influence on the deer population growth rate. Areas of recent timber harvest had 55% more forage biomass than older forests, but horizontal visibility did not differ, suggesting that timber harvest did not influence predation risk. Although proximity to roads did not affect the overall population growth rate, vehicle collisions caused a substantial proportion of deer mortalities, and reducing these collisions could be a win-win for deer and humans. The influence of apex predators and forage indicates a dual limitation by top-down and bottom-up factors in this highly human-modified system, suggesting that a reduction in apex predators would intensify density-dependent regulation of the deer population owing to limited forage availability.

Spring black bear harvest and predation pressure on moose calves in a multi‐predator system

AbstractThe moose (Alces alces; mooz in Anishinaabemowin, Ojibwe language will follow scientific names) is a vital subsistence food source to Anishinabe people of the midwestern United States and has recently declined in Minnesota, USA, with poor calf survival as a contributing factor. Predation is the primary cause of moose calf mortality and we explored whether calf predation rates could be reduced through management of a single predator in a multi‐predator system. Thus, we examined predation rates and causes of calf mortality before (2013–2015) and during (2016–2018) implementation of a spring black bear (Ursus americanus; makwa) harvest season, using baits to attract black bears, on the Grand Portage Indian Reservation, Minnesota, where black bears and gray wolves (Canis lupus; ma'iinganag) are the primary predators of moose calves. We validated our early study findings for the 5 years following the initial study with intermittent spring bear hunting seasons, from 2019–2023. The spring bear hunt was canceled because of a pandemic lockdown in 2020, resumed 2021, and was closed in 2022 and 2023. Black bear harvest prior to adding a spring bear hunting season was 0.038 bears harvested/km2 from 2012–2015, whereas after initiating a spring hunting season (2016–2018) it was 17% higher at 0.046 bears/km2. We observed significantly lower bear predation (by 68%) in association with spring bear management and no compensatory change in the level of wolf predation. The validation years strengthened our findings that spring bear hunting seasons reduce moose calf predation rates, with an overall 68% lower proportion of bear predation on moose calves in the years when spring bear hunts were held. Mean proportion of calf predation attributed to bears was 4.9 times higher (30% vs. 6%) in the years when a spring bear hunt was not held. Despite an increasing wolf density during the study period, we did not observe a compensatory increase in wolf predation during spring bear hunt years. The results of this work suggest that the addition of a spring bear hunt, during a time when moose calves are most vulnerable to bear predation, has the potential to increase moose calf survival even in the presence of wolves.

Prey size reflected in tooth wear: a comparison of two wolf populations from Sweden and Alaska.

Ingesta leaves distinct patterns on mammalian teeth during mastication. However, an unresolved challenge is how to include intraspecific variability into dietary reconstruction and the biomechanical aspects of chewing. Two extant populations of the grey wolf (Canis lupus), one from Alaska and one from Sweden, were analysed with consideration to intraspecific dietary variability related to prey size depending on geographical origin, sex and individual age as well as tooth function. Occlusal enamel facets of the upper fourth premolars, first molars and the second lower molar were analysed via three-dimensional surface texture analysis. The Swedish wolves displayed facets characterized by higher peaks and deeper, more voluminous dales, featuring an overall rougher surface than the wolves from Alaska. Compared to females, the Swedish male wolves had a slightly larger dale area and hill volume on their facets. Upper fourth premolars are smoother and had higher values in texture direction compared to upper first molars. The upper first molars were rougher than the occluding lower second molars and were characterized by larger and deeper dales. We find evidence supporting intraspecific dietary segregation, and antagonistic asymmetry in occlusal wear signatures. The data offer new insights into the roles of apex predators like the grey wolf.

Somatometric Characteristics of the Grey Wolf (Canis lupus) in Bulgaria

Grey wolf (Canis lupus Linnaeus 1758) is the largest representative of the Canidae family in Bulgaria. It inhabits almost the whole territory, most frequently the mountainous and hilly areas. The grey wolf is a hunting species in the country. Despite this, there is little information in the literature on its size. The aim of this work was to provide the first systematized data of the somatometric characteristics (weight – P; total body length – L; length of the body to the base of the tail – Lb; tail length – Lo and height at the withers – Hb) of the grey wolf in Bulgaria and to establish what is the degree of their sexual dimorphism. We found that the adult males are 24.3% heavier than the females (t=4.46; p<0.001). They also have a longer body by 8.8% (t = 3.66; p<0.001) and a greater height by 6.5% (U=546; p=0.005). The smallest difference (6.2%) between the two sexes was observed in tail length (U=629.5; p=0.03). The results of our study support the clear sexual dimorphism established in other wolf populations. Further studies are needed to link the wolf ’s body size data with prey type data in the respective areas for more clarity. This will be beneficial both for better wolf population management and for the accumulation of a database.  

Moose in wolf diets across northeastern Minnesota

AbstractThe moose (Alces alces; mooz in Anishinaabemowin, Ojibwe language) population has recently declined in Minnesota, USA, and gray wolf (Canis lupus; ma'iingan) predation is likely a contributing factor. We analyzed diet composition of gray wolves in northeastern Minnesota during 2011–2013 to evaluate the importance of moose as prey and seasonal and regional variations in wolf diet. We identified frequency of occurrence of prey items and biomass consumed in 1,000 wolf scats collected on and adjacent to the Grand Portage Indian Reservation and Voyageurs National Park and within the 1854 Ceded Territory (greater northeastern Minnesota). White‐tailed deer (Odocoileus virginianus; waawaashkeshiwag [plural]), moose, and beaver (Castor canadensis; amikwag [plural]) composed the majority of wolf diet, with moose as the primary prey in Grand Portage (35–54% of diet by biomass across seasons) and deer the primary prey in the 1854 Ceded Territory (46–62%) and Voyageurs (63–79%). Relative importance of prey species differed by study area and season. Moose calves were an important prey item in spring in the 1854 Ceded Territory (12% of diet by biomass) but not in Grand Portage or Voyageurs. Although calves were not a majority of wolf diet by biomass, many calves were preyed upon by wolves (30% of calves born each year in Grand Portage), thus affecting recruitment in a declining moose population. Deer fawns composed 12% of wolf diet in spring and 10% in summer in Grand Portage and 19% in summer in Voyageurs. Beaver composed 16% of wolf diet by biomass in spring and 14% in summer in Grand Portage and composed 22% of wolf diet in spring and 30% in summer in Voyageurs. At most prey densities, moose were preferred and deer avoided in Grand Portage and the 1854 Ceded Territory and beaver were preferred in Voyageurs. Our results can be used in conjunction with predation and prey studies to evaluate the effect of wolves on prey populations.

Domestication constrains the ability of dogs to convey emotions via facial expressions in comparison to their wolf ancestors

Dogs (Canis lupus familiaris) are the domestically bred descendant of wolves (Canis lupus). However, selective breeding has profoundly altered facial morphologies of dogs compared to their wolf ancestors. We demonstrate that these morphological differences limit the abilities of dogs to successfully produce the same affective facial expressions as wolves. We decoded facial movements of captive wolves during social interactions involving nine separate affective states. We used linear discriminant analyses to predict affective states based on combinations of facial movements. The resulting confusion matrix demonstrates that specific combinations of facial movements predict nine distinct affective states in wolves; the first assessment of this many affective facial expressions in wolves. However, comparative analyses with kennelled rescue dogs revealed reduced ability to predict affective states. Critically, there was a very low predictive power for specific affective states, with confusion occurring between negative and positive states, such as Friendly and Fear. We show that the varying facial morphologies of dogs (specifically non-wolf-like morphologies) limit their ability to produce the same range of affective facial expressions as wolves. Confusion among positive and negative states could be detrimental to human–dog interactions, although our analyses also suggest dogs likely use vocalisations to compensate for limitations in facial communication.

Lifetime reproductive characteristics of gray wolves

Abstract Female and male cooperative breeders can use different strategies to maximize reproduction and fitness over their lifetimes. Answering questions about fitness in cooperative breeders requires long-term studies as well as complete data on group composition and size which can be exceedingly difficult to obtain. Using a long-term genetic data set of complete group pedigrees, I asked how lifetime reproductive characteristics of female and male gray wolves (Canis lupus) differed. I predicted that genetic relatedness to helpers would be higher for females than males due to philopatric behavior of female wolves, group size would be similar between the sexes, females would inherit breeding positions from within groups more often than males due to differences in dispersal strategies between the sexes, males would have more lifetime mates and produce more young than females because of polygamy, and females would breed for more years than males due to the likelihood that females would still breed (with a new partner) after a mate died or was expelled from the group. I documented complete lifetime breeding histories for 11 male and 18 female wolves in Idaho, United States, 2008 to 2018. Genetic relatedness to helpers, group size, number of mates, pups, and years breeding did not differ between the sexes. Females, however, inherited breeding positions within groups far more often than males. Individuals who secured breeding positions generally reproduced for 2 seasons and commonly had more than 1 partner during their lifetimes if they were able to maintain their breeding position longer. Direct fitness varied greatly within female and male breeding wolves.

Distribution and potential habitat of the Vulnerable Himalayan wolf Canis lupus chanco in Bhutan

Abstract The rare Himalayan wolf Canis lupus chanco is categorized as Vulnerable on the IUCN Red List, and there is limited knowledge of its ecology and distribution. In Bhutan, the Himalayan wolf is one of the least known carnivores. Our aims in this study were to map the current distribution of the wolf in Bhutan and to identify potential habitats within the country. We compiled 32 records of wolf presence from camera-trap surveys and, using a maximum entropy approach, we estimated a potential habitat of 2,431 km2, comprising c. 6.3% of Bhutan. However, wolf presence was localized and non-continuous. We recommend a detailed fine-scale habitat analysis in areas of potential habitat and genetic studies to investigate population structure. Knowledge of these matters will provide insights regarding connectivity and facilitate the development of conservation strategies for this threatened wolf.

Craniometrical sexual dimorphism of the grey wolf (Canis lupus, Canidae, Carnivora) in Bulgaria

Cranial sexual dimorphism in grey wolf (Canis lupus Linnaeus, 1758) from Bulgaria has not been studied so far. Skulls from adult individuals from across the country were studied. Thirty-six parameters (35 craniodental measurements and a volumetric one) of each skull were measured. The Bulgarian wolf population exhibits weak craniodental sexual dimorphism, which is male-biased, as is typical in canids with a monogamous social system. However, the sexual dimorphism is more pronounced than in the studied more northern populations.

Behavioral trade-offs and multitasking by elk in relation to predation risk from Mexican gray wolves.

Predator non-consumptive effects (NCE) can alter prey foraging time and habitat use, potentially reducing fitness. Prey can mitigate NCEs by increasing vigilance, chewing-vigilance synchronization, and spatiotemporal avoidance of predators. We quantified the relationship between Mexican wolf (Canis lupus baileyi) predation risk and elk (Cervus canadensis) behavior. We conducted behavioral observations on adult female elk and developed predation risk indices using GPS collar data from Mexican wolves, locations of elk killed by wolves, and landscape covariates. We compared a priori models to determine the best predictors of adult female behavior and multitasking. Metrics that quantified both spatial and temporal predation risk were the most predictive. Vigilance was positively associated with increased predation risk. The effect of predation risk on foraging and resting differed across diurnal periods. During midday when wolf activity was lower, the probability of foraging increased while resting decreased in high-risk areas. During crepuscular periods when elk and wolves were most active, increased predation risk was associated with increased vigilance and slight decreases in foraging. Our results suggest elk are temporally avoiding predation risk from Mexican wolves by trading resting for foraging, a trade-off often not evaluated in behavioral studies. Probability of multitasking depended on canopy openness and an interaction between maternal period and predation risk; multitasking decreased prior to parturition and increased post parturition in high-risk areas. Openness was inversely related to multitasking. These results suggest adult female elk are altering the type of vigilance used depending on resource availability/quality, current energetic needs, and predation risk. Our results highlight potentially important, but often-excluded behaviors and trade-offs prey species may use to reduce the indirect effects of predation and contribute additional context to our understanding of predator-prey dynamics.

The Taurus Mountains, the Hotspot of Western Palearctic Biodiversity, Is in Danger: Marble Quarries Affect Wildlife

The Taurus Mountains in the Mediterranean Coastal Basin, considered a biodiversity hotspot, have a rich biodiversity in the Western Palearctic. The number of marble quarries in the Taurus Mountains has dramatically expanded over the past ten years. The objectives of this study are to (i) determine the impacts of quarrying on wildlife and (ii) determine the potential impacts of quarrying on the future of Taurus. A total of 57,547 photos and video images were analyzed on 5447 photo-trap days in two areas, the marble quarries and the control areas. Using 97 randomly selected marble quarries, the area they cover and their annual growth rates were determined. The most commonly seen animals were the wolf (Canis lupus), fox (Vulpes vulpes), lynx (Lynx lynx), and wild boar (Sus scrofa) in the control area, and the jackal (Canis aureus) and hare (Lepus europaeus) in the marble quarries (p < 0.001). Additionally, we found a significant positive correlation between the distance from the geographical center of the marble quarries and the number of dates of wolf, fox and wild boar sightings, with a significant negative correlation for hares (p < 0.05). A positive correlation was found between the area of marble quarries and the duration of operation (R = 0.89, p < 0.00). The waste from quarries, which makes up 79.7% of the total land used for this purpose, is the greatest cause of habitat degradation. According to calculations, even if no new marble quarries are built as of right now, 7.14% of the Taurus Mountains may have disappeared by the year 2027, and by the year 2032, 8.25% of the Taurus ecosystems may have disappeared completely. The Taurus Mountains, a center of Western Palearctic biodiversity, are being threatened by marble quarries. This study advances our knowledge of how marble quarries may affect wildlife. New strategies must be developed as soon as possible to protect the Taurus Mountains, the hotspot of the Mediterranean basin.

Effects of lethal management on gray wolf pack persistence and reproduction in Wisconsin, USA.

Direct human-caused mortality accounts for about half of all large mammal mortality in North America. For social species like gray wolves (Canis lupus), the death of pack members can disrupt pack structure and cause pack dissolution, and mortality of breeding adults or wolves during reproduction and pup-rearing can decrease pup recruitment. We estimated minimum and maximum probability of wolf pack persistence in Wisconsin, USA, during biological years (15 April-14 April) 2011-2019 and evaluated the influence of pack size and legal harvest mortality on pack persistence during 2012-2014. Harvests comprised 75-161 mortalities within 194 monitored packs during 2012-2014, with 56-74% of packs having no wolves harvested each year. As an index of reproduction during 2013-2019, we also estimated the proportion of packs where pups responded to howl surveys. We evaluated the influence of pack size, legal harvest, and agency removal on reproduction during 2013-2015. Annual maximum pack persistence probability was uniformly high (0.95-1.00), and annual minimum pack persistence probability ranged from 0.86-0.98 with a possible decline during years of harvest. Reproduction was similar in years following harvest and agency removal (2013-2015, pup response = 0.27-0.40), and years without harvest or agency removal the year prior (2016-2019, pup response = 0.28-0.66). Pack size had a positive effect on pack persistence and reproduction. Total number of wolf mortalities and number of adult male and females removed did not influence pack persistence or reproduction. We suggest that low per-pack mortality, timing of harvest and agency removal, and harvest characteristics during 2012-2014 supported stable pack persistence and reproduction.

Dietary analysis of wolf (Canis lupus) – a comparison of markers and methods

Metabarcoding is emerging as an alternative to morphological methods in noninvasive carnivore diet analysis based on scats. A number of metabarcoding markers have been developed but their comparative performance to recover DNA from scats remains mostly untested. We tested three markers covering a wide taxonomic range of prey items and compared them with the results of a morphological analysis. Morphological and genetic methods performed comparably regarding the identity of detected prey species, but the number of identified species varied strongly between markers. Only one, 12S-V5, amplified successfully in all samples and proved to be robust and reliable when working with the highly degraded DNA obtained from scats.

Spatial overlap of gray wolves and ungulate prey changes seasonally corresponding to prey migration

BackgroundPrey are more vulnerable during migration due to decreased familiarity with their surroundings and spatially concentrated movements. Predators may respond to increased prey vulnerability by shifting their ranges to match prey. Moose (Alces alces) and white-tailed deer (Odocoileus virginianus) are primary gray wolf (Canis lupus) prey and important subsistence species for Indigenous communities. We hypothesized wolves would increase use of ungulate migration corridors during migrations and predicted wolf distributions would overlap primary available prey.MethodsWe examined seasonal gray wolf, moose, and white-tailed deer movements on and near the Grand Portage Indian Reservation, Minnesota, USA. We analyzed GPS collar data during 2012–2021 using Brownian bridge movement models (BBMM) in Migration Mapper and mechanistic range shift analysis (MRSA) to estimate individual- and population-level occurrence distributions and determine the status and timing of range shifts. We estimated proportional overlap of wolf distributions with moose and deer distributions and tested for differences among seasons, prey populations, and wolf sex and pack affiliations.ResultsWe identified a single migration corridor through which white-tailed deer synchronously departed in April and returned in October–November. Gray wolf distributions overlapped the deer migration corridor similarly year-round, but wolves altered within-range distributions seasonally corresponding to prey distributions. Seasonal wolf distributions had the greatest overlap with deer during fall migration (10 October–28 November) and greatest overlap with moose during summer (3 May–9 October).ConclusionsGray wolves did not increase their use of the white-tailed deer migration corridor but altered distributions within their territories in response to seasonal prey distributions. Greater overlap of wolves and white-tailed deer in fall may be due to greater predation success facilitated by asynchronous deer migration movements. Greater summer overlap between wolves and moose may be linked to moose calf vulnerability, American beaver (Castor canadensis) co-occurrence, and reduced deer abundance associated with migration. Our results suggest increases in predation pressure on deer in fall and moose in summer, which can inform Indigenous conservation efforts. We observed seasonal plasticity of wolf distributions suggestive of prey switching; that wolves did not exhibit migratory coupling was likely due to spatial constraints resulting from territoriality.

A panel of tetranucleotide STR markers as an alternative approach to forensic DNA identification of wolf and dog.

Commercial panels of microsatellite (STR) loci are focused on the use of DNA of the domestic dog (Canis lupus familiaris) and are often inapplicable for genotyping the DNA of the gray wolf (Canis lupus lupus). We propose a CPlex test system, including one hexa- and 12 tetranucleotide autosomal STR loci, as well as two sex loci, that is equally efficient in DNA identification of biological samples of the wolf and the dog. Analysis of molecular variance between samples revealed significant differentiation values (FST = 0.0784, p < 0.001), which allows to use the panel to differentiate wolf and dog samples. Population subdivision coefficients (θ-values) were calculated for each of the 13 STR loci of the developed test system. It was shown that the values of the genotype frequency for dogs and wolves, without and with considering the θ-value, differ by three orders of magnitude (for dogs 8.9 × 10-16 and 2.1 × 10-14 and for wolves 1.9 × 10-15 and 4.5 × 10-14, respectively). The use of population subdivision coefficients will allow to identify the most reliable results of an expert identification study and the power of exclusion provided by the STR loci of the CPlex test system makes it possible to achieve a reliable level of evidence in forensic DNA analysis of both wolves and dogs. The test system has been validated for use in forensic identification of the dog and wolf based on biological traces found at crime scenes, as well as for individual identification and establishing biological relationship of animals of these species.

Suspected True Hermaphroditism in a Free-ranging Gray Wolf (Canis lupus) in Interior Alaska, USA.

Hermaphroditism or intersexuality occurs endogenously in most mammal species. We document the behavior and physical and anatomic appearance of a suspected true hermaphroditic free-ranging wild gray wolf (Canis lupus) in Denali National Park and Preserve, Alaska, USA.

Native prey, not landscape change or novel prey, drive cougar (Puma concolor) distribution at a boreal forest range edge.

Many large carnivores, despite widespread habitat alteration, are rebounding in parts of their former ranges after decades of persecution and exploitation. Cougars (Puma concolor) are apex predator with their remaining northern core range constricted to mountain landscapes and areas of western North America; however, cougar populations have recently started rebounding in several locations across North America, including northward in boreal forest landscapes. A camera-trap survey of multiple landscapes across Alberta, Canada, delineated a range edge; within this region, we deployed an array of 47 camera traps in a random stratified design across a landscape spanning a gradient of anthropogenic development relative to the predicted expansion front. We completed multiple hypotheses in an information-theoretic framework to determine if cougar occurrence is best explained by natural land cover features, anthropogenic development features, or competitor and prey activity. We predicted that anthropogenic development features from resource extraction and invading white-tailed deer (Odocoileus virgianius) explain cougar distribution at this boreal range edge. Counter to our predictions, the relative activity of native prey, predominantly snowshoe hare (Lepus americanus), was the best predictor of cougar occurrence at this range edge. Small-bodied prey items are particularly important for female and sub-adult cougars and may support breeding individuals in the northeast boreal forest. Also, counter to our predictions, there was not a strong relationship detected between cougar occurrence and gray wolf (Canis lupus) activity at this range edge. However, further investigation is recommended as the possibility of cougar expansion into areas of the multi-prey boreal system, where wolves have recently been controlled, could have negative consequences for conservation goals in this region (e.g. the recovery of woodland caribou [Rangifer tarandus caribou]). Our study highlights the need to monitor contemporary distributions to inform conservation management objectives as large carnivores recover across North America.

Cognitive and Intellectual Behavior of a Wolf (Canis lupus L.): Barriers Preventing Consuming the Elk (Alces alces) Population by a Predator

Cognitive and Intellectual Behavior of a Wolf (Canis lupus L.): Barriers Preventing Consuming the Elk (Alces alces) Population by a Predator

Nurturing orphaned Indian Grey Wolf at Machia Biological Park, Jodhpur, India

The Indian Grey Wolf Canis lupus pallipes is an iconic species of grassland in India. It is a Schedule 1 species according to the Wildlife Protection Act, 1972 and CITES Appendix 1 species. In the absence of parental care, five wolf pups were rescued from Baavarla on 26 November 2020 and brought into Machia Biological Park for hand rearing. In the absence of colostrum, a milk replacer formula (PetLac) was provided to the pups. The pups were kept at an ambient room temperature of 101°F as an adjustment to their natural surroundings. Gripe water, Vitamin A, and multivitamins were administered orally. The temperature and body growth parameters were measured and recorded weekly. Rabies vaccine was administered in the 26th week from the date of rescue. The diet of the wolf was gradually changed and from the 37th week, raw meat was included in their diet. Antibiotics Ciplox-TZ (ciprofloxacin and tinidazole tablet) Brand-Cipla and SPORLAC-DS (lactic acid bacillus tablets 120M) Brand-sanzyme Ltd. were given periodically to restore the gut fauna of the pups.