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Pre-implantation embryos of Chinese hamster. I. Incidence of karyotype anomalies in 226 control embryos.

Karyotyes were determined in 226 pre-implantation embryos (4--8-cell stages) of Chinese hamster. The study was carried out under controlled natural breeding conditions, without superovulation and with the embryos developing in their mothers. A total of 5.3% karyotypically abnormal embryos were found. Over half, 3.1%, were due to ploidy mutations, 5 cases of triploidy and 2 cases of haploidy. Only 0.9% genome mutations were present, consisting of one autosomal trisomy and one autosomal monosomy. Structural aberrations were found in 1.8%, half of these probably due to a balanced maternal aberration and the rest appearing the mosaic condition only. These results are compared with the scarce body of mammalian data from the literature. Compared with the situation in man, the spontaneous aberration rates in the Chinese hamster and other experimental mammals are extremely low. This may be due, in part, to optimal timing of copulation in respect to estrus and ovulation prevailing in these animals but not in man. The low spontaneous aberration rate in the reported system is a valuable asset for purposes of mutagen testing.

Animals↗

The origins of supraspinal projections to the cervical and lumbar spinal cord at different stages of development in the gray short-tailed Brazilian opossum, Monodelphis domestica.

We have used the retrograde transport of Fast blue (FB) to study the origins of supraspinal projections to the lumbar and cervical spinal cord at different stages of development in the Brazilian, short-tailed opossum, Monodelphis domestica. Monodelphis was chosen for study because its young are born in a very immature state, 14-15 days after copulation, making it possible to manipulate its nervous system in an embryonic state without intra-uterine surgery. When injections of FB were made into the lumbar cord at postnatal day (PD) 1, neurons were labeled within several areas of the reticular formation (the retroambiguus nucleus, the ventral and dorsal reticular nuclei of the medulla, the gigantocellular reticular nucleus, the lateral paragigantocellular reticular nucleus, and the pontine reticular nucleus), the presumptive coeruleus complex, and the lateral vestibular nucleus. In many cases, labeled neurons were also found within the caudal raphe and the presumptive interstitial nucleus of the medial longitudinal fasciculus. The results of immunocytochemical studies provided evidence for catecholaminergic and serotoninergic neurons in the brainstem at PD1 and for axons of both phenotypes in the spinal cord. By PD3, labeled neurons were found within the ventral gigantocellular and ventral pontine nuclei of the reticular formation, the spinal trigeminal nucleus, and the presumptive paraventricular nucleus of the hypothalamus. When injections were made at PD4, neurons were also labeled within the medial and inferior vestibular nuclei, the red nucleus, the mesencephalic nucleus of the trigeminal nerve, the presumptive nucleus of Edinger-Westphal and the lateral hypothalamus. By at least PD7, the pattern of supraspinal labeling was similar to that obtained at older ages and in the adult animal. When FB was injected into the cervical cord at PD1, neurons were labeled in all of the areas labeled by lumbar injections at the same age and in larger numbers. In addition, labeled neurons were found within the ventral gigantocellular and spinal trigeminal nuclei. When cervical injections were made at PD15, labeled neurons were found within the deep cerebellar nuclei and amygdala and by PD17 they were also present within the superior colliculus and cerebral cortex. In some cases, cortical labeling was present outside the areas labeled by comparable injections in adult animals.(ABSTRACT TRUNCATED AT 400 WORDS)

Amidines↗

The early development of major projections from caudal levels of the spinal cord to the brainstem and cerebellum in the gray short-tailed Brazilian opossum, Monodelphis domestica.

The Brazilian short-tailed opossum, Monodelphis domestica, is born 14-15 days after copulation and is available for experimentation at stages of development corresponding to those which occur in utero in placental mammals. In the present study, we took advantage of the opossum's embryology to study the development of projections from caudal levels of the spinal cord to the brainstem and cerebellum using axonal tracing methods. In all cases, a 2-3 day survival time was used for axonal transport. When injections of Fast blue (FB) were made into caudal levels of the thoracic cord at postnatal day (PD) 1 or 2, axonal labeling could not be identified at supraspinal levels. When injections were made at PD3, however, labeled axons were found in the fasciculus gracilis at caudal medullary levels, within the ventrolateral medulla and pons, within an incipient inferior cerebellar peduncle, and within the cerebellar anlage. The dorsal root origin of at least some of the axons within the fasciculus gracilis was evidenced by the transganglionic transport of cholera toxin conjugated to horseradish peroxidase from the hindlimbs. After FB injections at PD7, a few labeled axons could be traced from the fasciculus gracilis into the nucleus gracilis and from the ventrolateral pathway to the inferior olive. Generally comparable results were obtained using wheat germ agglutinin conjugated to horseradish peroxidase (WGA-HRP). In cases injected with FB at PD9, the pattern of brainstem labeling was adult-like. Although labeled axons were present within the cerebellum of animals injected with FB on PD3, they were limited to the marginal zone. Axonal labeling was present within an identifiable internal granular layer in cases injected with either FB or WGA-HRP at PD16, and it appeared to be limited to specific bands which foreshadowed those seen at later stages of development and in the adult animal. In some cases, labeled axons were present within the molecular layer where they were not seen in the adult animal. Our results provide a timetable for the normal development of projections from caudal levels of the spinal cord to the brainstem and cerebellum in Monodelphis and show that such development occurs postnatally rather than prenatally, as in placental mammals.

Aging↗

Sexual motivation of the male rat during the oestrous cycle of the female rat.

Two experiments are described in which the sexual motivation of the male rat was investigated in relation to the reproductive state of a female rat. The male rat lived in an artificial habitat, the residential plus-maze, so that continuous registration of his movements was possible without disturbing the animal. It was found that a male rat showed a preference for a female rat in heat even though copulation was not possible. Such behaviour was also shown by the male rat towards an ovariectomized female that had been brought in heat by oestradiol benzoate with a subsequent injection of progesterone. A single injection with oestradiol benzoate given to the female rat could not induce this behaviour in the male rat. The nature of the signals that inform the male rat about the female condition has not yet been elucidated.

Animals↗

Midbrain lesions, dopamine and male sexual behavior.

Destruction of the medial preoptic area (MPOA) eliminates mating in male rats and this region is believed to play a major role in the control of male sexual behavior. Efferents from the MPOA pass through and/or terminate in 4 midbrain regions: the dorsolateral tegmentum (DLT), the central gray, and the A9 and A10 areas. The present study reports the effects of bilateral destruction of each of these midbrain regions on brain catecholamines and sexual behavior in male rats. DLT lesions eliminated mating, reproducing the effect of bilateral preoptic lesions. The sexual activity of males with central gray lesions was accelerated in the sense that the mounting rate for these males was significantly faster than for controls and lesioned males ejaculated more frequently and with shorter latencies than did controls. A9 lesions impaired mating--lesioned males mounted at a slower rate and ejaculated less frequently than controls. Males with A10 lesions took longer to re-initiate mating after an ejaculation than controls, but copulation per se (as reflected in mount rate, ejaculation frequency and latency to ejaculate) was not affected by A10 damage. Brain catecholamine levels were not affected by either DLT or central gray lesions. A9 lesions produced a significant depletion in neostriatal dopamine which was highly correlated with mount latency, mount rate, ejaculation latency and ejaculation frequency. A10 lesions produced a significant depletion of dopamine in the nucleus accumbens and cingulate cortex, but these effects were not significantly correlated with any measure of sexual behavior.

Animals↗

Structural and functional characterization of neuropeptides involved in the control of male mating behavior of Lymnaea stagnalis.

Mating as a male in the simultaneous hermaphrodite freshwater snail, Lymnaea stagnalis, comprises a series of complex behaviors that are a prelude to copulation. Copulatory behavior itself is assumed to be controlled by various types of peptidergic neurons as well as serotonergic cells. Here we report the primary structure of two peptides that were extracted from a cluster of neurons that innervates the penial complex and that is located in the anterior lobe of the right cerebral ganglion. The sequences of the peptides were determined as: Ala-Pro-Gly-Trp-amide and Ser-Gly-Ser-Asp-Tyr-Cys-Glu-Thr-Leu-Lys-Glu-Val-Ala-Asp-Glu-Tyr-Ile-Leu- Leu- Ser-Tyr-Lys-Ile-Glu-Glu-Gln-Arg-Ala-Ala-Asp-Cys-Gly-Gly-Glu-Pro-Pro-Asn- Ser- Gln(amide), respectively. The longer peptide is a homodimer. Both peptides are processed from the recently identified Ala-Pro-Gly-Trp-amide prohormone, which is expressed in the neurons of the anterior lobe of the right cerebral ganglion. Ala-Pro-Gly-Trp-amide could also be recovered from the penial complex. This peptide, when applied in vitro, inhibits the contractions of the penis retractor muscles evoked by serotonin in a dose-dependent fashion.

Amino Acid Sequence↗

Restoration of sexual function and fertility by fetal hypothalamic transplant in impotent aged male rats.

The purpose of this study was to observe whether fetal hypothalamic transplant can restore the neuroendocrine and reproductive function in impotent aged male rats. Eighteen to 20 month old impotent male rats received an anterior hypothalamus removed from a 17-19 day old fetus and placed into the anterior third ventricle. Controls were either without surgery (UC) or grafted with cerebral cortex (CG). Before and 2 to 3 months after transplantation, blood samples were collected from the aged rats for testosterone and LH measurement. Before and one to two months after transplantation, each hypothalamic grafted animal (HG) or control rat was put overnight into a cage which contained four, 10 to 12 week old proestrous female rats. Vaginal smear of each female was monitored early the next morning. Sperm seen in the vaginal smear was regarded as copulation and ejaculation. The test was repeated twice, one week apart, and the higher score represented the sexual function and fertility of the males. Seven of 10 HG males restored their sexual function, impregnated 9 females and fathered 106 pups. None of 7 UC restored their reproductive function and only one of 4 CG males impregnated one female which delivered 6 pups. Serum testosterone, LH and pituitary LH in the HG rats, which showed restoration of reproductive function, were significantly higher than those of the controls (UC and CG). These results indicate that the fetal hypothalamic grafts can survive and develop in the brain of impotent aged male rats and restore neuroendorince and reproductive function in senescent rats.

Aging↗

Aging-induced decrements in neuropeptide Y: the retention of ejaculatory behavior is associated with site-selective differences.

Neuropeptide Y (NPY) has been implicated in the control of reproductive and cardiovascular function. We observed an age-related decrease in the number of males copulating to ejaculation and a moderate systolic hypertension in middle-aged (16- to 17-month-old) rats. NPY content was examined in microdissected brain nuclei in 5 groups of rats: 2 groups of young rats, 1 heterosexually naive and the other ejaculating in 3 successive mating test; 3 groups of middle-aged rats, 1 heterosexually naive, 1 group that had extensive sexual experience but failed to ejaculate in tests at 16.5 months of age, and the third continuing to ejaculate at 16.5 months of age. NPY levels were found to vary depending on the brain area, the age of the animals, and the maintenance of ejaculatory behavior. In sexually naive middle-aged males, NPY levels were uniformly lower than in younger males. There were no differences in NPY levels of young animals, regardless of sexual experience. In the medial preoptic area, the group that retained ejaculatory behavior through 16.5 months of age, had higher levels of NPY than those observed in young sexually experienced rats. In sexually experienced rats that were no longer ejaculating at 16.5 months of age levels were lower than all other groups except the sexually naive middle-aged group. In the hypothalamic arcuate nucleus, levels were equivalent in the young groups and in the middle-aged rats that retained ejaculatory behavior, being greater than in the middle-aged rats that were no longer ejaculating or were sexually naive.(ABSTRACT TRUNCATED AT 250 WORDS)

Aging↗

Reproduction in Fischer-344 rats exposed to methyl chloride by inhalation for two generations.

Male and female Fischer-344 rats were exposed to methyl chloride by inhalation (0, 150, 475, or 1500 ppm, 6 hr/day, 5 days/week, 40 males and 80 females per group). The only treatment-related clinical signs were a 10 to 20% body weight gain depression (BWGD) in both males and females exposed to 1500 ppm at all weekly weighings after 2 weeks of exposure and a 5-7% BWGD in 475-ppm exposed animals after Day 57. After 10 weeks the exposure schedule was changed to 6 hr/day, 7 days/week and each male was mated to two exposed females. The mating period was ended after 2 weeks, at which point 10 males/group were necropsied. The only treatment-related lesions found were severe bilateral testicular degeneration (10/10) and granulomas in the epididymis (3/10) in the 1500-ppm males. The remaining 30 males per group were then removed from exposure and mated during a 2-week period with 60 unexposed females. The exposed females were continued on exposure from the start of mating to Postnatal Day 28 (6 hr/day, 7 days/week). The females were not exposed from Gestation Day 18 to Postnatal Day 4, and the pups were never directly exposed prior to weaning. There were no significant differences between groups in the number of exposed or unexposed females that mated, as evidenced by copulation plugs. No litters were born to exposed or unexposed females mated to the 1500-ppm males. There was no significant difference in the number of litters produced by the 150-ppm groups when compared to the control groups. Fewer litters were born in the 475-ppm groups than in the control groups. No differences in litter size, sex ratio, pup viability, or pup growth were found among the 475-ppm, 150-ppm, or control F0 groups. When bred 10 weeks after the cessation of exposures, 5 to 20 1500-ppm F0 males had regained the ability to sire normal litters. The same number of 475-ppm F0 males proved as fertile (15/20) as control F0 males (13/20). After weaning, F1 pups from the 475-, 150-, and 0-ppm groups were exposed to the same concentrations of methyl chloride for 10 weeks and then mated. A trend toward decreased fertility was found in the 475-ppm F1 group.

Animals↗

Developmental toxicity of dimethylacetamide by inhalation in the rat.

Dimethylacetamide (DMAC) is a widely used industrial solvent. It has been reported to be teratogenic when given to rats by injection or following dermal application. Most of these studies employed large single doses and did not examine both the fetal and the maternal response. In this study, groups of pregnant Crl:CD rats were exposed to 32, 100, or 282 ppm DMAC by inhalation for 6 hr/day from Days 6 through 15 of gestation (day on which copulation plug was detected was termed Day 1G). A control group of chambered pregnant rats was exposed simultaneously to air only. All female rats were euthanized on Day 21G. At 282 ppm, both maternal weight gain during the exposure period and fetal weight were significantly decreased and accompanied by a significant dose-response trend. These effects were not seen in rats inhaling either 32 or 100 ppm. Fetal resorptions were not increased in any of the groups exposed to DMAC. Fetal incidences of external, visceral, or skeletal variations and malformations were similar between the test and control groups. Therefore, both fetal and maternal toxicity were noted at 282 ppm and the no-observed adverse-effect level under these experimental conditions was 100 ppm for both the dam and the conceptus. DMAC was not demonstrated to produce malformations in the rat fetus even at a level that was toxic to the dam.

Abnormalities, Drug-Induced↗

Effect of Prograf (FK506) on spermatogenesis in rats.

Prograf (FK506) was given to male mature rats in a daily subcutaneous dose of 1 or 3 mg/kg/day for 2 weeks to investigate its effect on spermatogenesis. Prograf dose-dependently sperm counts and motility, but did not affect testosterone level in the serum of rats. Histopathologically, there were no abnormal changes in the testis, seminal vesicle or prostate in any rats dosed with Prograf, but intra-ductal eosinophilic globules, probably degeneration of the sperm cells, were observed in the epididymis of the 3 mg/kg/day group. Sperm counts and motility returned to the control levels after stopping of the drug. The results indicate that Prograf decreased sperm counts and motility through direct action on the sperm in the epididymis, but not the production of sperm in the testis. Cyclosporine A (CsA) was used as the reference drug in the present study. Thirty mg/kg/day of CsA also decreased sperm counts and motility, and stopping of the drug led to the recovery of these changes. The males dosed with Prograf for 2 weeks were mated with non-dosed females to investigate its effect on the fertility potential of the males. Prograf did not affect copulation or fertility index, but a decrease in the number of live fetuses associated with implantation loss was observed in the 3 mg/kg/day group. The changes were considered to be due to the decrease of sperm counts and motility, although 1 mg/kg/day of Prograf, 5-10 times the clinical dose, did not affect any fertility parameters including implantation index.

Animals↗

Dopamine functions in appetitive and defensive behaviours.

The data reviewed here are compatible with the hypothesis that telencephalic dopamine activity is elicited by motivationally significant stimuli which in turn creates a neural state in which animals are more prepared to respond to significant stimuli in the environment. This analysis may be viewed as extensions of both the sensorimotor hypothesis, which depicts dopamine as potentiating the ability of stimuli to elicit responses (Clody and Carlton, 1980; Marshall et al., 1974; White, 1986) and of the incentive motivational hypothesis, which emphasizes the importance of dopamine in responding to stimuli that serve as signals of biologically significant events (Blackburn et al., 1989a; Crow, 1973; Mogenson and Phillips, 1976). In addition, we have sought to emphasize that not all responses are equally dependent upon the integrity of forebrain dopamine activity. Some responses, such as ingestion of standard foods by hungry animals, copulation, and escape, are relatively impervious to dopamine disruption. Further, once other behaviours, such as avoidance or appetitive operant responses, have been acquired, they can be maintained at an initially high rate despite perturbation of dopamine systems, although performance deteriorates with repeated testing. This analysis has emerged from the joint consideration of how both appetitive and defensive behaviours are influenced by dopamine antagonists, along with an examination of dopamine release during sequences of behaviour. The data reviewed suggest that dopamine is involved in fundamental psychological processes through which environmental stimuli come to exert control over certain aspects of behaviour. In the future, as knowledge in this field advances, there will have to be an integration of the literature on dopamine and motivation with the literature on dopamine and motor systems. We expect that dopamine release will be seen as a mechanism by which important environmental cues, of innate or learned significance, lead to a general enhancement of motor skeletal responses directed towards distal cues. We conclude with a caveat: Caution must be exercised when attempting to infer a general role of any neurotransmitter in motivated behaviour based on the study of a limited number of motivational systems. Although neurotransmitter pathways may figure prominently in the control of certain behaviours, it is incorrect to think of neurotransmitters as having a single role in behaviour. However, when comparative analyses reveal a common thread among different motivational systems, as is becoming apparent for the general role of mesotelencephalic dopamine pathways in behaviour, then the goal of generating coherent and comprehensive theory concerning a neurotransmitter's function in behaviour will begin to be realised.(ABSTRACT TRUNCATED AT 400 WORDS)

Aggression↗

Behavioral changes induced by GABA-receptor agonists in Lymnaea stagnalis L.

1. GABA, the GABAA receptor agonist, muscimol, and the GABAB receptor agonist, baclofen, were tested to study the involvement of the GABA neurotransmitter system in control of behaviour in the freshwater pulmonate snail, Lymnaea stagnalis L. Single injections of GABA (1-10 micrograms/ gbw) into the haemocoel of intact snails elicits a sequence of behavioural changes subsequently affecting feeding, locomotion, escape reactions, male mating behaviour and respiration. 2. Both muscimol and baclofen mimic distinct aspects of GABA action implying that the GABA action is mediated by both types of receptors. 3. The modulatory actions of GABA, muscimol, and baclofen on feeding were manifested as opening of the mouth and triggering of specific radular movements (e.g., protraction, retraction, rasping). 4. Baclofen (1-10 micrograms/gbw) evoked the full erection of the penis for a time long enough for natural copulation, while GABA itself caused only partial eversion of the preputium. Muscimol was less effective than GABA. The latency to penis eversion varied in a dose-dependent manner. The data emphasize the participation of baclofen-sensitive receptors in control of reproduction. 5. GABA and baclofen induced withdrawal of the head and anterior part of the foot accompanied by arrest of locomotion and respiration could be considered to be an escape reaction. 6. Muscimol (1-5 micrograms/gbw) failed to elicit withdrawal reactions but caused the loss of normal orientation and longitudinal contractions of the foot. 7. All the agents tested inhibited locomotion in a dose-dependent manner, for a substantial period of time. A decrease in total locomotor activity lead to the failure of animals to attach to the underlying surface, to have foot contractions and also to diminish locomotor velocity. 8. The majority of animals maintained the stereotyped complex reactions of respiration, but with restricted clockwise turning of the shell after drug treatment. 9. All the substances were shown to shift the background behavioural state characterised by high BSSs to a state similar to "rest" which was awarded a low BSS. It is concluded that the GABA neurotransmitter system can play an important role in activation and coordination of neuronal ensembles underlying behaviour and behaviour selection in Lymnaea stagnalis L.

Animals↗

The effects of beta-endorphin infusions into the amygdala on visual and olfactory sensory processing during sexual behaviour in the male rat.

Sexually experienced male rats infused bilaterally into the amygdala with 60 pmol beta-endorphin show decreased rate of precopulatory investigation of the female and delayed intromission latency, but copulation is left unaltered. Such males are still able to discriminate between the odours of bedding from receptive and unreceptive females, demonstrating that beta-endorphin does not impair the ability to detect sexually relevant odours. Preventing visual cues emitted by females during proceptive behaviour (by treating them with haloperidol) delayed intromission latency but had no effect on preintromission investigation. Intra-amygdaloid beta-endorphin exacerbated the effects of this treatment on the intromission latency. Inducing anosmia in males (by applying zinc sulphate solution to the olfactory mucosa) decreased their anogenital investigation and delayed their intromission latency. These effects were not enhanced by intra-amygdaloid beta-endorphin. Allowing males to investigate and initiate the first intromission prior to intra-amygdaloid infusion had no effects on subsequent intromissions. However, if following an intromission with one female and an infusion of beta-endorphin, the male was presented with an unfamiliar female then the effects of intra-amygdaloid beta-endorphin on investigation and intromission returned. These results suggest that beta-endorphin in the amygdala interferes with the processing of female-specific olfactory information. Without this processed information, classification of the female as a sexual stimulus may be impeded and thus sexual arousal delayed.

Amygdala↗

A thirteenth cranial nerve: the cloacal nerve.

The completion by vertebrates of micturition, defecation, and copulation via the cloaca or its derivatives is hypothesized to be best explained by the existence of a thirteenth cranial nerve, the cloacal nerve, which, similar to the facial and trigeminal nerves, functions as a mixed cranial nerve containing both general and special components.

Animals↗

Implication of testosterone metabolism in the control of the sexually dimorphic nucleus of the quail preoptic area.

In quail, testosterone (T) activates male copulation and affects the volume and cytoarchitectonic organization of the medial preoptic nucleus (POM). T metabolism (especially its aromatization) is critical for the production of these behavioral effects. We wondered whether T metabolism is also playing a role in the induction of the morphological changes in POM. We compared the effects of T and of its metabolites in this nucleus. To obtain an independent evaluation of the role played by aromatase, morphological effects of T associated or not with the aromatase inhibitor R76713 were also assessed. As previously observed, T increased the POM volume and the cross-sectional area of the neurons in the lateral part of the nucleus. The effects of T on the neurons in the lateral POM were mimicked in part by the combined treatment with estradiol and 5 alpha-dihydrotestosterone. They were also blocked by the aromatase inhibitor. This suggests that T aromatization plays a critical role in the mediation of the cytoarchitectonic effects of T. A specific role for androgens alone remains to be established.

Animals↗

Effects of mating status, sex ratio, and population density on longevity and offspring production of Cryptolestes ferrugineus (Stephens) (Coleoptera: Cucujidae).

The life span and reproductive rate of the rusty grain beetle, Cryptolestes ferrugineus (Stephens), were determined at different adult densities and sex ratios at 30 +/- 1 degree C, 75 +/- 5% relative humidity (RH), and with feed consisting of ground wheat plus wheat germ (4:1, w/w). The mean life spans of adult beetles were 32 weeks for adults individually isolated in separate vials, 22 to 25 weeks for adults of different strains kept one male and one female per vial, 24 weeks for a group of 30 virgin females together in one vial, 13 weeks for a group of 30 virgin males together in one vial, 14 weeks for groups with 10 males and 20 females per vial, 13 weeks for groups with 15 males and 15 females per vial, and 12 weeks for groups with 20 males and 10 females per vial. When kept one adult per vial, males and females had similar life spans; when kept one male and one female per vial, males lived longer; when kept in groups of 30 per vial females lived longer, except in the group which had a sex ratio of two males to one female; in this group life spans of males and females were the same. Females in separate vials with one male produced mean numbers of of offspring ranging from 389 for an inbred homozygous malathion-resistant strain to 514 for an outbred strain (genetically variable: GV). GV strain females in vials which had 10 males and 20 females produced a mean of 97 offspring, GV strain females in vials which had 15 males and 15 females produced a mean of 146 offspring, and GV strain females in vials which had 20 males and 10 females produced a mean of 216 offspring. The results of this study and similar studies on other insect species suggest that the life span of this insect is inversely related to the rate at which it expends energy and to injury incurred during copulation, and that oviposition rate decreases as the density of larvae and female adults increases.

Animals↗