New evidence for hybrid zones of forest and savanna elephants in Central and West Africa

The African elephant consists of forest and savanna subspecies. Both subspecies are highly endangered due to severe poaching and habitat loss, and knowledge of their population structure is vital to their conservation. Previous studies have demonstrated marked genetic and morphological differences between forest and savanna elephants, and despite extensive sampling, genetic evidence of hybridization between them has been restricted largely to a few hybrids in the Garamba region of northeastern Democratic Republic of Congo (DRC). Here, we present new genetic data on hybridization from previously unsampled areas of Africa. Novel statistical methods applied to these data identify 46 hybrid samples – many more than have been previously identified – only two of which are from the Garamba region. The remaining 44 are from three other geographically distinct locations: a major hybrid zone along the border of the DRC and Uganda, a second potential hybrid zone in Central African Republic and a smaller fraction of hybrids in the Pendjari–Arli complex of West Africa. Most of the hybrids show evidence of interbreeding over more than one generation, demonstrating that hybrids are fertile. Mitochondrial and Y chromosome data demonstrate that the hybridization is bidirectional, involving males and females from both subspecies. We hypothesize that the hybrid zones may have been facilitated by poaching and habitat modification. The localized geography and rarity of hybrid zones, their possible facilitation from human pressures, and the high divergence and genetic distinctness of forest and savanna elephants throughout their ranges, are consistent with calls for separate species classification.     

Forest elephant mitochondrial genomes reveal that elephantid diversification in Africa tracked climate transitions

Among elephants, the phylogeographic patterns of mitochondrial (mt) and nuclear markers are often incongruent. One hypothesis attributes this to sex differences in dispersal and in the variance of reproductive success. We tested this hypothesis by examining the coalescent dates of genetic markers within elephantid lineages, predicting that lower dispersal and lower variance in reproductive success among females would have increased mtDNA relative to nuclear coalescent dates. We sequenced the mitochondrial genomes of two forest elephants, aligning them to mitogenomes of African savanna and Asian elephants, and of woolly mammoths, including the most divergent mitogenomes within each lineage. Using fossil calibrations, the divergence between African elephant F and S clade mitochondrial genomes (originating in forest and savanna elephant lineages, respectively) was estimated as 5.5 Ma. We estimated that the (African) ancestor of the mammoth and Asian elephant lineages diverged 6.0 Ma, indicating that four elephantid lineages had differentiated in Africa by the Miocene–Pliocene transition, concurrent with drier climates. The coalescent date for forest elephant mtDNAs was c. 2.4 Ma, suggesting that the decrease in tropical forest cover during the Pleistocene isolated distinct African forest elephant lineages. For all elephantid lineages, the ratio of mtDNA to nuclear coalescent dates was much greater than 0.25. This is consistent with the expectation that sex differences in dispersal and in variance of reproductive success would have increased the effective population size of mtDNA relative to nuclear markers in elephantids, contributing to the persistence of incongruent mtDNA phylogeographic patterns.     

Estimating the defaecation rate of the African forest elephant (Loxodonta cyclotis) in Banyang-Mbo Wildlife Sanctuary, south-western Cameroon

Defaecation rate is an important parameter in the conversion of dung counts to elephant numbers, yet estimates of elephant defaecation rates are available for a few sites, especially rainforest sites. The Banyang‐Mbo Wildlife Sanctuary in south‐western Cameroon is no exception in this regard. We tracked a radio‐collared herd of four forest elephants, counted their fresh droppings in two 24‐h periods each month in 36 consecutive months. Our results show a mean defaecation rate of 15.9 [95% CI (12.2–19.7)] dung piles per elephant per day. We found significant monthly and inter‐annual variation in defaecation rates with implications for their predictability and forest elephant surveys. In particular, our results show that mean defaecation rates are higher for the wettest period of the year than the relatively dry months.     

African elephant age determination from teeth: validation from known individuals

We assess tooth‐based age criteria for African elephants developed by Laws in relation to known‐aged individuals in the Amboseli elephant population. Laws’s technique remains a robust and useful mechanism for age determination, although we suggest revisions to the oldest age categories. Blind age assignment to jaws of unknown sex using the Laws criteria resulted in misclassification of M4 and M5; measures overlapped too much to differentiate these teeth by sex. Sexes could be reliably distinguished after age 30 or XIX in tooth category by two measures: mandible thickness and width of the ascending ramus, but individuals of the same known age differed in tooth wear and progression rates. Such variation needs to be incorporated in the error assigned to tooth age categories. Ages at death of found jaws (n = 266) were similar to results of survival analysis from all demographic data (n = 2455), excluding calves whose jaws decompose because of weathering and scavengers. Jaw‐based models of age at death need correction for the inability to detect this early mortality, which artificially extends mean longevity by up to 6 years.     

Acoustic estimation of wildlife abundance: methodology for vocal mammals in forested habitats

Habitat loss and hunting pressure threaten mammal populations worldwide, generating critical time constraints on trend assessment. This study introduces a new survey method that samples continuously and non‐invasively over long time periods, obtaining estimates of abundance from vocalization rates. We present feasibility assessment methods for acoustic surveys and develop equations for estimating population size. As an illustration, we demonstrate the feasibility of acoustic surveys for African forest elephants (Loxodonta africana cyclotis). Visual surveys and vocalizations from a forest clearing in the Central African Republic were used to establish that low‐frequency elephant calling rate is a useful index of elephant numbers (linear regression P < 0.001, r_adj.² = 0.58). The effective sampling area was 3.22 km² per acoustic sensor, a dramatic increase in coverage over dung survey transects. These results support the use of acoustic surveys for estimating elephant abundance over large remote areas and in diverse habitats, using a distributed network of acoustic sensors. The abundance estimation methods presented can be applied in surveys of any species for which an acoustic abundance index and detection function have been established. This acoustic survey technique provides an opportunity to improve management and conservation of many acoustically‐active taxa whose populations are currently under‐monitored.     

Patterns of molecular genetic variation among African elephant populations

The highly threatened African elephants have recently been subdivided into two species, Loxodonta africana (savannah or bush elephant) and L. cyclotis (forest elephant) based on morphological and molecular studies. A molecular genetic assessment of 16 microsatellite loci across 20 populations (189 individuals) affirms species level genetic differentiation and provides robust genotypic assessment of species affiliation. Savannah elephant populations show modest levels of phylogeographic subdivision based on composite microsatellite genotype, an indication of recent population isolation and restricted gene flow between locales. The savannah elephants show significantly lower genetic diversity than forest elephants, probably reflecting a founder effect in the recent history of the savannah species.     

Vegetation changes during a 36-year period in northern Chobe National Park, Botswana

Changes in vegetation cover in northern Chobe National Park (Botswana) were assessed using aerial photographs from 1962, 1985 and 1998, with subsequent ground proofing. In addition, cumulative browsing by elephants and the occurrence of fire scars were recorded on random vegetation sites within shrubland (n = 20) and mixed woodland (n = 20). Coverage of woodland vegetation decreased from 60% to 30% between 1962 and 1998, while shrubland vegetation increased from 5% to 33% during the same period. During the study period, woodland has gradually retreated away from the river front. While riparian forest covered a continuous area along the riverfront in 1962, only fragments were left in 1998. We found a significant decrease in browse use with increasing distance to the Chobe river for Combretum apiculatum, Combretum elaeagnoides, Combretum mossambicense and other woody plants combined (all P < 0.0001). The occurrence of fire (P < 0.0001) and basal area (P < 0.0001) were positively related to distance to the river. Elephant browsing occurred on >70% of available stems within 2 km from the river, while less than 20% of the trees had fire scars in the same zone. Beyond 7 km from the river, elephant browsing was reduced to >50% of available stems, while more than 50% of the trees had fire scars. The density of any of the shrubs was not related to distance to the river neither within shrubland (all P > 0.05) nor within mixed woodlands (all P > 0.05).     

亚洲象繁殖生物学特征探讨

运用全天观察和记录法,对一只连续繁殖两胎的雌性亚洲象在发情、交配、妊娠、分娩、产后行为及幼象行为等方面的表现做了详细观察和记录,并结合文献资料进一步探讨了亚洲象繁殖生物学特征。结果表明：①采用适当的食物、药物、人工授精等人工辅助措施有助于亚洲象的繁殖;②母象妊娠体征明显,中后期乳头间距增大并能挤出粘性液体,可作为判断母象怀孕的重要依据;③加强对母、幼象的护理利于健康和成活;④幼象日吮乳次数逐日减少;⑤幼象1周岁断奶是可行的;⑥不同胎次的妊娠征兆、怀孕期、幼象排大便等特征有较大差异。     

Enzyme‐immunoassay for the measurement of luteinizing hormone in the serum of African elephants (Loxodonta africana)

Circulating patterns of progesterone and luteinizing hormone (LH) in the elephant have been well characterized, and routine monitoring of these hormones is now viewed as a valuable tool for making informed decisions about the reproductive management of elephants in captivity. Currently, LH monitoring in elephants is done with radio‐immunoassays (RIAs); unfortunately, the use of radioactive materials in RIAs limits their application to institutions with laboratory facilities equipped for the storage and disposal of radioactive waste. Enzyme‐immunoassays (EIAs) offer an inexpensive and more zoo‐friendly alternative to RIA. This work reports on an EIA capable of quantifying circulating LH in African elephants. The EIA employs a biotin label and microtiter plates coated with goat anti‐mouse gamma globulin. LH surges in African elephants (n=3) increased fivefold over baseline concentrations (1.00±0.1 ng/ml vs. 0.2±0.1 ng/ml) and occurred 19.3±0.2 days apart. Ovulatory LH surges were associated with an increase in serum progestogens from 4.8±0.4 ng/ml to 11.7±0.4 ng/ml. The ability to quantify reproductive hormones in elephants via EIA is an important step in the process of making endocrine monitoring more accessible to zoos housing these species. Zoo Biol 21:403–408, 2002. © 2002 Wiley‐Liss, Inc.     

Molecular Characterization of Pro-Inflammatory Cytokines Interleukin-1β and Interleukin-8 in Asian Elephant (Elephas maximus)

Interleukin (IL)-1β and IL-8 are pro-inflammatory cytokines produced primarily by monocytes and macrophages in response to a variety of microbial and nonmicrobial agents. As yet, no molecular data have been reported for IL-1β and IL-8 of the Asian elephant. In the present study, we have cloned and sequenced the cDNA encoding IL-1β and IL-8 of the Asian elephant. The open reading frame (ORF) of Asian elephant IL-1β is 789 bp in length, encoded a propeptide of 263 amino acid polypeptide. The predicted protein revealed the presence of IL-1 family signature motif and an ICE cut site. Whereas, IL-8 contained 321 bp of open reading frame. Interestingly, the predicted protein sequence of 106 aa, contains an ELR motif immediately upstream of the CQC residues, common in all vertebrate IL-8 molecules. Identity levels of the nucleic acid and deduced amino acid sequences of Asian elephant IL-1β ranged from 68.48 (Squirrel monkey) to 98.57% (African elephant), and 57.78 (Sheep) to 98.47% (African elephant), respectively, whereas that of IL-8 ranged from 72.9% (Human) to 87.8% (African elephant), and 63.2 (human, gorilla, chimpanzee) to 74.5% (African elephant, buffalo), respectively. The phylogenetic analysis based on deduced amino acid sequenced showed that the Asian elephant IL-1β and IL-8 were most closely related to African elephant. Molecular characterization of these two cytokines, IL-1β and IL-8, in Asian elephant provides fundamental information necessary to progress the study of functional immune responses in this animal and gives the potential to use them to manipulate the immune response as recombinant proteins.     

Urinary androstenedione and luteinizing hormone concentrations during musth in a mature African elephant

Musth has not been well documented in captive African elephants. A 37-year-old African bull elephant in the Kansas City Zoological Park was observed during periods of behavioral musth and non-musth. Androstenedione and luteinizing hormone (LH) concentrations in urine were measured by radioimmunoassay. Urinary androstenedione and LH levels were significantly higher in musth urine than in non-musth samples. A positive correlation (P > 0.001) existed between urinary LH and androstenedione concentrations. These results indicate that musth can occur in a zoo-maintained African elephant and that urinary androgen levels are elevated during musth, possibly as the result of LH stimulation of testicular steroidogenesis.     

Concentrations of progesterone,testosterone and estradiol-17β in the serum during the estrous cycle of Asian elephants (Elephas maximus)

The levels of progesterone, testosterone and estradiol-17β in serum samples from two female Asian elephants were measured for the period of 32 months from February 1987 to September 1989. Serum samples were collected weekly from unanesthetized elephants. Each elephant showed eight ovarian cycles in 32 months. Ovarian cycles, characterized by changes in concentrations of serum progesterone, averaged 16.8 ± 0.6 (mean ± SEM. n = 14) weeks in length. The changes in concentrations of testosterone in the serum showed a similar pattern to those of progesterone with a striking increase noted during the luteal phase. The highest levels of serum estradiol-17β were noted when progesterone levels showed low basal values. These results suggest that estradiol-17β may be an index of follicular maturation during the estrous cycle in Asian elephants, and that the ovaries of Asian elephants may produce testosterone in the luteal phase.     

Early functional maturity of captive male northern elephant seals (Mirounga angustirostris)

A male northern elephant seal (Mirounga angustirostris) that was conceived in captivity was stillborn February 14, 1981, at Mystic Marinelife Aquarium in Mystic, Connecticut. Either of two males in the exhibit could have sired the pup. Both were only 3 yr at the time of conception.     

中国亚洲象取食植物种类统计与分析

2016年1月至2017年12月,分别采用样线法和跟踪调查法对中国境内分布的野生亚洲象和云南西双版纳亚洲象种源繁育与救助中心的10头圈养亚洲象放养时取食植物进行调查,经分类鉴定,共收集到亚洲象取食植物111种,分属29目、42科、77属,其中新增亚洲象取食植物57种,分属于21目、32科、44属。与文献中记载的中国亚洲象植物性食物进行了汇总,统计得到新的中国亚洲象植物性食物名录,共计32目、62科、162属、240种。多样性分析结果显示,G-F多样性指数为0.84,表明中国野生及圈养亚洲象取食的植物具有较高的多样性,这些植物涉及蕨类植物门、裸子植物门和被子植物门。其中,圈养亚洲象采食植物种类共84种,西双版纳境内圈养亚洲象以禾本科(Gramineae)的甘蔗(Saccharum spp.)、玉米(Zea mays)、马唐(Digitaria spp.)、象草(Pennisetum purpureum)、粽叶芦(Thysanolaena latifolia)等为主要食物,而国内其它地方(北京动物园、广州动物园和昆明动物园)圈养的亚洲象主要饲喂高粱属(Sorghum)的高粱(S.bicolor)或苏丹草(S.sudanense)等。因此,在将来开展野生亚洲象栖息地保护、恢复及食物源基地项目规划与建设时应规避农作物和经济作物,优选亚洲象喜食、速生、生物量大的土著物种,如野芭蕉、棕叶芦、董棕、构树、重阳木、中平树、马唐属及竹类等植物,按照不同季节进行套种,为亚洲象提供更多可口食物。对于圈养亚洲象,需要设计足够面积的食物源基地供放养,补充食物种类,增加运动量,增强体质,提高其生活质量。     

Social factors influence ovarian acyclicity in captive African elephants (Loxodonta africana)

Nearly one-third of reproductive age African elephants in North America that are hormonally monitored fail to exhibit estrous cycle activity, which exacerbates the nonsustainability of the captive population. Three surveys were distributed to facilities housing female African elephants to determine how social and environmental variables contribute to cyclicity problems. Forty-six facilities returned all three surveys providing information on 90% of the SSP population and 106 elephants (64 cycling, 27 noncycling and 15 undetermined). Logistic analyses found that some physiological and social history variables were related to ovarian acyclicity. Females more likely to be acyclic had a larger body mass index and had resided longer at a facility with the same herdmates. Results suggest that controlling the weight of an elephant might be a first step to helping mitigate estrous cycle problems. Data further show that transferring females among facilities has no major impact on ovarian activity. Last, social status appears to impact cyclicity status; at 19 of 21 facilities that housed both cycling and noncycling elephants, the dominant female was acyclic. Further studies on how social and environmental dynamics affect hormone levels in free-living, cycling elephants are needed to determine whether acyclicity is strictly a captivity-related phenomenon. Zoo Biol 28:1–15, 2009. © 2008 Wiley-Liss, Inc.     

Six new polymorphic microsatellite loci isolated and characterized from the African savannah elephant genome

The African savannah elephant (Loxodonta africana) is a ‘keystone’ species that plays a vital role in regulating the dynamics of both plant and animal communities and yet it is endangered and its numbers have been reduced to approximately 500 000 across their entire continental range. Molecular genetic markers are important tools for providing genetic information useful in formulating effective management and conservation strategies for the surviving elephant populations. We describe the isolation and characterization of six new polymorphic microsatellite markers in the African savannah elephant and demonstrate that these loci can be PCR (polymerase chain reaction)‐multiplexed, a desirable attribute that saves costs in large‐scale microsatellite screening.