Risk Management Proposal  
Semen and Embryos from Horses  (Equidae ) 
 
 
 
 
 
 
 
 
 
December  2015   
 
 
 
Disclaimer  
 
This risk management proposal does not constitute, and should not be regarded as, legal advice. While every 
effort has been made to ensure the information in this document is accurate, the Ministry for Primary Industries 
does not accept any responsibility or liability whatsoever for any error of fact, omis sion, interpretation or opinion 
that may be present, however it may have occurred.  
 
Requests for further copies should be directed to:  
 
Ministry for Primary Industries  
Regulation & Assurance  
Animal Imports  
PO Box 2526  
WELLINGTON 6140  
 
Email:   animal.imports@mpi.govt.nz  
Telephone:          04 890 0134  
Facsimile:             04 894 0733  
 
© Crown Copyright - Ministry for Primary Industries   
i Contents  Page 
 
1 Purpose  1 
2 Background  1 
3 Objective  2 
4 General Requirements  3 
5 Recommendations for Identified Risk Organisms  3 
5.1 Borna disease virus  3 
5.2 Burkholderia mallei (glanders)  4 
5.3 Equine herpesvirus -1 (EHV -1) 5 
5.4 Equine infectious anaemia virus (EIA)  6 
5.5 Equine arteritis virus (Equine viral arteritis, EVA)  8 
5.6 Leptospira  species  10 
5.7 Salmonella  species  11 
5.8 Taylorella species (Contagious equine metritis, CEM)  12 
  
Ministry for Primary Industries   Risk Management Proposal for Semen and Embryos from Horses ( Equidae )  1 1 Purpose  
The purpose of this document is to:  
 Show how options for the management of risk organisms in semen and embryos from horses have been 
assessed  
 Provide recommendations for import requirements  for semen and embryos from horses . 
 
The import health standard (IHS)  for semen and embryos from horses from specified countries  is developed 
under Section 24 of the Biosecurity Act 1993.  
 
For a detailed risk assessment of the identified hazard s, refer to the Import Risk Analysis: Equine germplasm 
from Australia, Canada, the European Union and the USA  (22 December 2009) .  A copy can be viewed at the 
following link: http://www.biosecurity.govt.nz/files/regs/imports/risk/equine -germplasm -ra.pdf  
 
2 Background  
Equine semen and embryos are considered risk commodities, with the poten tial to harbour exotic viral, bacterial 
and protozoal diseases.  In December 2009, the Ministry for Agriculture and Forestry  (now the Ministry for 
Primary Industries, MPI) completed the Import Risk Analysis: Equine germplasm from Australia, Canada, the 
European Union and the USA.  This import risk analysis ( 2009 IRA) forms the basis of the risk management 
measures in the proposed IHS.  
 
Norway requested inclusion into the new IHS for semen and embryos from horses . A comparison of the 
Norwegian animal health situation (WAHID 2014)  with that of EU member countries  has supported the decision to 
allow imports of equine semen and embryos  from Norway.  Similarly, Switzerland was included after comparing 
the animal health situation with that of other EU countries. Se e Table 1 for more information.  
 
The following IHSs for individual countries or groups of countries will be consolidated into one IHS which will 
contain generic import req uirements for semen and embryos:   
 
 Import Health Standard for Horse Semen from Austra lia, 10 November 2004  
 Import Health Standard for Horse Semen from Canada , 8 August 2005  
 Import Health Standard for Equine Semen from the European Union , 1 August 2007  
 Import Health Standard for Horse Semen from the USA , 12 February 2009 . 
 
In addition to the IHS, a guidance document for equine semen and embryos will be issued. This document will 
include model veterinary certificates.  
Ministry for Primary Industries   Risk Management Proposal for Semen and Embryos from Horses ( Equidae )  1 Table 1: Organisms of concern in semen and embryos of horses.  
Hazard  Transmissible 
in semen/ 
embryos  Presen t in 
Australia?  Present 
in EU?  Present 
in 
Norway?  Present in 
Switzerland?  Present in 
North 
America?  Transmitted 
only by 
insect or 
helminth 
vectors♥  Preliminary 
hazard for 
2009 IRA  
Viruses     
African horse 
sickness virus  Yes%/Yes%  No*  No*  No* No* No*  Yes@  No  
Vesicular stomatitis 
virus  Yes/Yes”  No*  No*  No* No* Yes*  Yes@  No  
Venezuelan equine 
encephalomyelitis 
virus  Yes”/Yes”  No*  No*  No* No* Not since 
1971*  Yes@  No  
Eastern equine 
encephalomyelitis 
virus  No/No  No*  No*  No* No* Yes*  Yes@  No  
Western equine 
encephalomyelitis 
virus  No/No  No*  No*  No* Not in 2013*  Yes*  Yes@  No  
Equine infectious 
anaemia virus  Yes/Yes  Yes*  Yes*  Not since 
1975*  Not in 2013, 
last report 
unknown*  Yes*  Mechanical 
insect vectors 
also in semen  Yes  
Equine influenza 
virus  No/No  No1 Yes*  Yes 
(2008)*  Yes (2009)*  Yes*  No@  Yes  
Equine herpesvirus -1 
(exotic strains)  Yes/No  Yes*  Yes*  Yes 
(2006)*  Yes*  Yes*  No@  Yes  
Equine arteritis virus  Yes/No  Yes*  Yes*  Yes*  Yes*  Yes*  No@  Yes  
Horse pox viruses  No/No  No*  No*  No* No No*  No@  No  
Japanese 
encephalitis virus  No/No  No*2 No*  No* No* No*  Yes@  No  
West Nile virus  No/No  No**  Yes**  No* No* Yes*  Yes@  No  
Rabies virus  No/No  
(uncertainty)  No*  Yes*  Yes?*  Last rep in 
2003*  Yes*  No@  Yes  
Borna disease virus  No/Yes  No**  Yes**  Yes** Yes No**  No@  Yes  
Equine encephalosis 
virus  No/unknown  No**  No**  No* No No@  Yes@  No  
Louping ill virus  No/unknown  No**  Yes**  Yes Yes No3 Yes@  No  
Hendra virus  Yes/Yes  Yes**  No**  No No No**  No@  Yes  
Getah virus  No/unknown  No**  No**  No No No**  Yes@ No  
      
Bacteria          
Bacillus anthracis  Yes/Yes  Yes*  Yes*  Yes 
(1993)*  Yes (1997)*  Yes*  No@  Yes  
Leptospira spp.  Yes/Yes  Yes*  Yes*  Yes  Yes*  Yes*  No@  Yes  
Taylorella 
equigenitalis  
Taylorella 
asinigenitalis  Yes/No  No*  Yes*  Yes Not in 2013* 
(last rep in 
2008)  Yes*  No@  Yes  
Burkholderia mallei  No/No No*  No*  Last 
reported 
1889  Last reported 
1939  No*  No@  No  
Burkholderia 
pseudomallei  No/Yes  Yes*  No*  No* No No4 No@  Yes  
Salmonella spp.  Yes/Yes  Yes*  Yes*  Yes Not in 2013  Yes*  No@  Yes  
      
Other organisms          
Ehrlichia risticii and 
Ehrlichia equi  No/No  No**  Yes**  Not in 
2013  Not in 2013  Yes  Yes@  No  
Histoplasma 
capsulatum var. 
farciminosum  No/No  No*  No*  No* No No*  No*  Yes#  
      
Other organisms 
from IRA 2000           
2  Risk Management Proposal for Equine Semen and Embryos  Ministry for Primary Industries   
* Country status as reported on the  World Animal Health Information Database (OIE 2013 ).  
** Disease distribution discussed in the MAF Import risk analysis: horses and horse semen (MAF 2000a) and cross -referenced in the textbook Veterinary 
Medicine (10th Ed) (Radostits et al 2007).  
% found in semen but no transmission evidence - To date there have been no known outbreaks due to the use o f infected semen, ova or embryos.  
@ As reported by the appropriate authors and in the appropriate sections In: Infectious Diseases of Livestock (Authors -various 2004).  
# Although not known to occur in the relevant countries it is hard to be certain as the  organism may be found in soil mud etc.  
♥ The organisms marked “yes” in column 8 are transmitted only by arthropod vectors or in one case a helminth vector ( Ehrlichia risticii ). These organisms 
are not transmissible via germplasm and therefore they are no t considered to be hazards in the commodity.  
 
3 Objective  
The objective is to manage, to an appropriate level of protection, the biosecurity risks for equine semen and 
embryos. Measures will align with the OIE Code  when possible.  
 
The biosecurity risks associated with the importation of equine semen and embryos have been identified in the 
2009 IRA.  Of the potential hazards identified in equine semen and embryos, risk management measures are 
justified for the following:  
 
Viral  
Equin e arteritis virus, OIE listed disease; unwanted, notifiable organism  
Equine herpesvirus -1 (exotic strains), OIE listed disease  
Equine infectious anaemia virus, OIE listed; unwanted, notifiable organism  
 
Bacterial  
Leptospir a spp (exotic serovars), other ex otic organisms  
Taylorella  equigentialis , unwanted, notifiable organism . 
 
 
Borna disease and equine salmonellosis were identified as potential hazards, but no measures for these diseases 
were warranted when import ing semen and embryos of horses.  
 Bovine brucellosis  
(B. abortus)  Yes/No  No* Some 
countries*  No Last in 1996*  No (Can), 
Yes (USA)*  No Yes 
Lyme disease  No/No  Yes Yes Yes?  Yes?  Yes Yes No 
Q fever  Yes/No  Yes*  Yes*  No Yes Yes*  Yes@  No 
Equine protozoal 
myeloencephalitis 
(Sarcocystis 
neurona ) No/No  Yes?  No No No Yes Yes No 
Dourine  Yes/No  No* Italy*  No Not in 2013*  No* No Yes 
Equine 
piroplasmosis   No/No  Not in 
2013*  Some 
countries*  No Yes*  Yes*  Yes No 
Nipah  Yes/unknown  No* No* No No* No* Yes No 
Surra  No/No  No* Not in 
2013*  No No* No* Yes No 
     
Ministry for Primary Industries   Risk Management Proposal for Semen and Embryos from Horses ( Equidae )  3 4 General Requirements  
The general requirements section of the IHS includes risk management measures for semen or embryo donor s, 
regardless of the country’s disease freedom claims. Where possible, t hese requirements align with the 
recommendations of the Code  and the International Embryo Transfer Society ( IETS) .   
 
The Code  provides methods to prevent germplasm from being contaminated with risk organisms during 
collection, processing, storage, and transport.  Although the Code 's methods for embryos are not specific to  
horses , they have been adapted for horses  in this document .  In the case of embryo collection, the Code  
describes three phases of risk: the health status of the herd or country of origin, the processes followed during 
collection of embryos, and the testing of embryos  collected or surveillance of the donors. It is recommended that 
the embryo  collection team should be approved by the Competent Authority. The team’s practices, as well as the 
processing and storage of embryos  and any test samples, should be in accordance with the Code  where 
applicable.  
  
If testing before collection is permitted, the donor shou ld be required to be isolated from other animals not of 
equivalent tested health status until the end of germplasm collection. On the day of collection, the health status of 
each donor should be confirmed to be free of clinical evidence of infectious disea ses transmissible in germplasm 
by the approved veterinarian. This should include examination (inspection, palpation) of the external reproductive 
organs. Additionally, if risk is managed by ensuring absence of disease from the donor and establishment for a  
specific amount of time after collection, the germplasm should be stored for the amount of time specified under 
each disease before export.  All germplasm should be stored in sealed,  tamper -proof,  labelled straws or 
containers.  
 
The IETS  Manual  provides guidance regarding washing and verification of the intact zona pellucida (intact and 
free of adherent material) after washing. These are important steps in processing to ensure that viruses will not 
be transmitted with the embryo. Micro -manipulat ion should only be permitted after washing and verification of the 
zona pellucida for maximum assurance that the embryo is free of virus.  
5 Recommendations for Identified Risk Organisms  
5.1 Borna disease virus 
5.1.1 Options for management of risk presented in th e 2009 IRA for equine germplasm   
Option 1  
i. Both male and female donors could come from countries certified by the veterinary authority as free 
from the disease; or  
ii. In countries where the disease does occur, and in which the disease is notifiable, animals co uld be 
certified as having been resident for the previous 3 months on a property on which the disease has 
not occurred during the previous 12 months.  
NB. this option is similar to the requirements in the current IHS for the importation of horses.  
Option 2  
Aliquots of semen or a sample of embryos and embryo washing solution from each batch of germplasm 
could be tested by RT -PCR for Borna disease virus RNA, with negative results.  
NB. This test is not validated for this purpose but could be justified on the grounds of probable high 
sensitivity. However, it is unlikely to be available in most testing laboratories.  
Option 3  
Aliquots of semen or a sample of embryos and embryo washing  fluid from each batch of germplasm could be 
cultured on cell cultures derived from embryonic rabbit or rat brain with negative results  
NB. This method is unlikely to be available in many (possibly most) testing laboratories.   
4  Risk Management Proposal for Equine Semen and Embryos  Ministry for Primary Industries  Option 4  
Aliquots of semen and a sample of each batch of embryos could be tested by intracerebral inoculation of 
rabbits, with negative results.  
NB. This test may be unacceptable on animal ethics grounds and unlikely to be available in many 
laboratories.  
5.1.2 Discussion  
Borna disease v irus is listed on the Unwanted Organisms R egister as an exotic organism. It is not an OIE 
listed disease.  
 
The requirements for Borna disease in the current IHS for horses ( HORANIIC.GEN, 2014 ) are as follows, 
simila r to Option 1:  
 
i. horses were kept since birth or for at least the past 90 days in a free country; or  
ii. horses were kept since birth or for at least the past 90 days on premises in which no case has been 
reported during the past 12 months.  
 
There are no measures for Borna  virus in the current IHSs for equine semen.  Borna disease is not OIE -
listed and is not important to trade. It occurs only sporadically in horses and sheep, and exceptionally in a 
variety of animals in parts of Germany. The epidemiology is not fully under stood, but current understanding is 
that there is reasonable evidence that this disease is spread by direct contact or fomites. There is no 
evidence that it is transmitted venereally or by traded germplasm. This goes for all species. Accordingly, any 
measu res for Borna disease are difficult to scientifically justify.  
 
5.1.3 Recommendation  
It is recommended no measures are put in place for Borna disease.  
 
5.2 Burkholderia mallei (glanders)  
5.2.1 Options for management of risk presented in the 2009 IRA for equine germplasm : 
 
Glanders was not identified as a hazard in the 2009 IRA for equine germplasm, and no measures were 
presented  in that document.  
5.2.2 Discussion  
In 2015, the OIE sought to amend the Code  chapter on glanders due to the disease re -emerging. 
Recommended measures w ere proposed for semen and embryos as follows:  
 
Recommendations  for the importation of equine semen  
Veterinary  Authorities  of importing countries  should require the presentation of an 
international veterinary certificate  attesting that : 
1. the donor males animals:  
a) showed no clinical signs of glanders on the day of collection;  
b) were examined clinically for signs of orchitis, with negative results . 
2. the semen was collected, processed and stored in accordance with the relevant  
recommendations in Chapter  4.5. and Articles 4.6.5. to 4.6.7.  
Recommendations for the importation of in vivo derived equine embryos   
Ministry for Primary Industries   Risk Management Proposal for Semen and Embryos from Horses ( Equidae )  5 Veterinary Authorities  of importing countries  should require the presentation of an 
international veterinary certificate  attesting that:  
1. the donor females showed no clinical signs of glanders on the day of collection  
2. the embryos were collected, processed and stored in accordance with the relevant 
recommendations in Chapters  4.7. and 4.9.;  
3. semen used to fertilise the oocytes complies with t he recommendations in the 
Article  recommending measures for the importation of equine semen.  
 
In 2015, MPI made a submission to the OIE that measures for germplasm are not warranted based on 
insufficient scientific basis to require such restrictions on either embryos or semen. The ad hoc  Group report 
that supports the inclusion of these articles in the Code  cites a single publication to justify the application of 
these measures, namely Khan et al. (2013) Glanders in animals: A review on epidemiology, clinical 
presentation, diagnosis and countermeasures. Transboundary and Emerging Diseases , 60, 204 -221. The ad 
hoc Group report summarises this review as stating that a large percentage of infected  equines had orchitis 
and therefore concluded that “it cannot be stated with any certainty that semen cannot transmit B. mallei  
infection”, and this same argument (orchitis) is used to justify the imposition of measures for the international 
trade in equin e embryos.  
The epidemiology section of the Khan et al. review paper cited makes no reference to the transmission of B. 
mallei  through equine germplasm although it does cite Saqib (2009) as describing 31/69 horses with glanders 
as having orchitis. Saqib (20 09) is a PhD thesis from the University of Faisalabad, Pakistan. The literature 
review of that thesis describes transmission of B. mallei  by ingestion or inhalation but makes no reference to 
venereal transmission (pp 20 -21). Although the thesis does descri be orchitis in a number of horses with 
glanders, the section of the thesis (pp 93 -94) suggests that this is actually the cutaneous form of glanders and 
is associated with contaminated bedding.  
The OIE’s Handbook on Import Risk Analysis for Animals and Ani mal Products  states that “It is not acceptable 
to simply conclude that, because there is significant uncertainty, measures will be based on a precautionary 
approach. The rationale for selecting measures must be made apparent”. In this case, there is no evi dence 
to suggest that B. mallei  is likely to be transmitted through the international trade in equine germplasm and 
the precautionary approach adopted by the inclusion of these articles is inconsistent with OIE guidance.  
Freedom from clinical signs on the day of collection and collection, processing and storage according to the 
relevant Code  recommendations are part of the general requirements to be met by semen and embryos 
from horses imported into New Zealand. Compliance of the semen used to produce the e mbryos with the 
OIE Code  is already a general requirement for the importation of embryos from horses. The only 
requirement that is an addition is the specific mentioning of freedom from clinical signs of orchitis upon 
clinical examination (palpation of the  testes). This requirement was added to the IHS under the semen 
donor requirements . 
5.2.3 Recommendation  
No specific requirements for managing glanders in semen and embryos from horses are necessary if the 
semen donors meet the general requirements.  
 
5.3 Equine her pesvirus -1 (EHV -1) 
5.3.1 Options for management of risk presented in the 2009 IRA for equine germplasm:  
 
For Semen  
Option 1  
Semen could be imported provided that donor stallions complied with the Code  requirement for live horses.  
Option 2   
6  Risk Management Proposal for Equine Semen and Embryos  Ministry for Primary Industries  Semen could be imported provided each batch of semen was tested by a RT -PCR test for the 
neuropathogenic mutant strain of EHV -1 with negative results.  
 
For Embryos  
Under the assumption that trypsin washing will be effective in removing any EHV -1 from equ ine embryos, 
embryos could be imported provided trypsin washing is used in addition to the existing I nternational Embryo 
Transfer Society  requirements for equine embryos.  
 
5.3.2 Discussion  
 
Although EHV -1 and EHV -4 are present in New Zealand, measures are recom mended to prevent the 
introduction of exotic, pathogeni c strains of EHV -1.  EHV  has been reported in  both equine semen and 
embryos and no country approved for export of semen and/or embryos to New Zealand is free of EHV -1. 
 
Trypsin may have an adverse effect on embryo viability. The current  IHS for horses (2013) requires that the 
horses meet the measures recommended in the OIE  Code . As imported horses pose a greater  risk of 
introducing EHV  than semen and embryos, the risk associated with imported semen and embryos should be 
effectively managed when derived from donors that have complied with the OIE Code  requirements for live 
horses. Furthermore, EHV -1 is endemic in New Zealand and hence the import measures imposed on 
germplasm should not be more stringe nt than those imposed on live horses.  
 
Current OIE Code  recommendations for the importation of equines  
Veterinary Authorities  of importing countries  should require the presentation of an 
international veterinary certificate  attesting that the animals : 
1. showed no clinical sign of equine herpes virus type 1 infection  (abortigenic and 
paralytic forms) on the day of shipment and during the 21  days prior to shipment;  
 
2. were kept for the 21  days prior to shipment in an establishment  where no case  of 
equine herpes virus type  1 infection  (abortigenic and paralytic forms), was reported 
during that period.  
5.3.3 Recommendation  
 
Option 1, d onor animals must meet the OIE Code recommendations for managing EHV -1 in horses . 
 
5.4 Equine infectious anaemia virus (EIA)  
5.4.1 Options for management of risk presented in the 2009 IRA for equine germplasm:  
 
For Semen  
Option 1  
Semen could be imported provided that:  
 
i. donors show no clinical sign of EIA on the day of semen collection and for the 60 days after 
semen collection: and  
ii. no case of EIA has been associated with any premises where the animals were kept during the 
3 months prior to and the 60 days after semen co llection.  
  
Ministry for Primary Industries   Risk Management Proposal for Semen and Embryos from Horses ( Equidae )  7 Option 2  
Semen could be imported provided that:  
i. donors show no clinical sign of EIA on the day of semen collection; and  
ii. no case of EIA has been associated with any premises where the donors were kept during the 3 
months prior to collection; and  
iii. Donors are subjected to an AGID test or ELISA for EIAV antibody not less than 21 days after 
entry onto the semen collection centre  with a negative result.  
 
 (Risk analysis subsequently recommended that the timing of the test be modified to not less than 30 days 
after entering the semen collection centre, to allow time for antibody development.)  
Option 3  
Semen could be imported provided that:  
 
i. donors show no clinical sign of EIA on the day of semen collection and for the 60 days after 
semen collection; and  
ii. no case of EIA has been associated with any premises where the animals were kept during the 
3 month s prior to and the 60 days after semen collection; and  
iii. donors are subjected to an AGID test or ELISA for EIAV antibody 30 -60 days after semen 
collection with negative results.  
 
NB. In most cases antibody can be detected by the AGID within 45 days of infection however, although rare , 
the period between exposure and antibody detection  can be as long as 60 days.  
 
For Embryos  
Option 1  
Embryos could be imported provided that:  
i. both male and female donors show no clinical sign of EIA on the day of semen collection and for 
the 60 days after germplasm collection: and  
ii. no case of EIA has been associated with any premises where the donor animals were kept during 
the 3 months prior to  and the 60 days after semen collection.  
Option 2  
Embryos could be imported provided that:  
i. both male and female donors show no clinical sign of EIA on the day of germplasm collections 
and for the 60 days after germplasm collection; and  
ii. no case of EIA h as been associated with any premises where the animals were kept during the 
3 months prior to and the 60 days after germplasm collection; and  
iii. donors are subjected to an AGID test or ELISA for EIAV antibody 30 -60 days after germplasm 
collection with negati ve results.  
5.4.2 Discussion  
EIA is an OIE listed disease and the virus is an unwanted, notifiable organism in New Zealand. The virus has 
been shown to be transmitted by semen. There is uncertainty regarding the risk of transmission via embryos. 
There are OIE Code  requirements for live horses, but none for semen and embryos.  
The 201 4 IHS for horses requires that horses meet the OIE Code , with samples for testing collected 
in pre -export isolation or in the 21 days prior to export if PEI is not required.  Extrapolating the Code  
recommendations for live horses to semen and embryos, testing could take place during the 30 
days prior to collection.  
 
Current OIE Code  recommendations for the importation of horses   
8  Risk Management Proposal for Equine Semen and Embryos  Ministry for Primary Industries  Veterinary Authorities  of importing countries  should require the presentation of an 
international veterinary certificate  attesting that:  
1. the animals  showed no clinical sign of equine infectious anaemia (EIA) on the day of 
shipment and during the 48  hours prior to shipment; and  
 
2. no case of EIA has been associated with any premises where the animals  were kept 
during the three  months prior to shipment; and  
 
3. if imported on a permanent basis, t he animals  were subjected to a diagnostic test for 
EIA with negative results on blood samples collected during the 30  days prior to 
shipment; or  
 
4. if imported on  a temporary basis, the animals  were subjected to a diagnostic test for 
EIA with negative results on blood samples collected during the 90  days prior to 
shipmen t. 
5.4.3 Recommendation  
The measures in the IHS to align  with current OIE Code recommendations for managing EIA in horse s. 
Extrapolated to germplasm donors, this means the following:  
(1) Donors showed no clinical sign of EIA on the day of each collection; and  
a) Donors were kept on premises where no case of EIA has been reported during the 90 days prior to 
each collection; and  
(1) Donors were subjected to a test listed in the MPI document: MPI Approved Diagnostic Tests, Vaccines, 
Treatments and Post -arrival Testing La boratories for Animal Import Health Standards (MPI -STD-TVTL) , 
not less than 30 days prior to collection with negative results.  
 
5.5 Equine arteritis virus (Equine viral arteritis, EVA)  
5.5.1 Options for management of risk presented in the 2009 IRA for equine germpla sm  
For Semen  
Donor stallions should conform to the recommendations of Article 12.10.4 of the Code . However, the Code  
recommendations are complex and contain a number of different options.  
For Embryos  
Option 1  
Embryos could be imported provided that:  
i. male donors meet the requirements in the relevant Articles  of the Code ; and  
ii. female donors have been vaccinated with an approved vaccine according to the manufacturer’s 
recommendations, at least 4 weeks before collection of embryos.  
Option 2  
Embryos could be imported provided that:  
i. male donors meet the requirements in the relevant Article s of the Code ; and  
ii. female donors are subjected to a serological test 1 week before and 3 weeks after collection of 
embryos. Female donors would be suitable donors if both serological tests are negative or if 
positive titres are stable or declining.   
Ministry for Primary Industries   Risk Management Proposal for Semen and Embryos from Horses ( Equidae )  9 5.5.2 Discussion  
EVA is an OIE listed disease and the virus is listed as an unwanted, notifiable organism in New Zealand. In 
2014 New Zealand was recognised by the OIE as free from EVA. T he virus is known to be shed in semen 
and stallions can remain carriers of the virus. The OIE Code  recommendations for semen will cover this risk.  
 
Transmission through embryos is unknown, and mares do not remain carriers of t he virus.  Option s 1 & 2 for 
female donors are similar to the OIE Code  recommendations for the importation of equids other than 
uncastrated males, with the timing  modified for embryo collection . However,  since the 2009 IRA was written, 
the OIE Code has recommended measures for trade in equine embryos.  
Current OIE Code  recommendations for the importation of equine semen  
Veterinary Authorities  of importing countries  should require the presentation of an 
international veterinary certificate  attesting that the donors were kept for the 28  days prior 
to semen collection in an establishment  where no equid has shown any clinical sign of EVA 
during that period and showed no clinical sign of EVA on the day of semen collection; and  
1. were subjected between six and nine  months of age to a test for EVA:  
2. EITHER:  
a) with a negative result,  
OR 
b) with a positive result, followed at least 14 days later by a second test showing a 
stable or decreasing titre;  
c) and were immediately vaccinated against EVA and regularly revaccinated in 
accordance with the recommendations of the manufacturer; or  
3. were isola ted and not earlier than seven  days of commencing isolation were subjected to 
a test for EVA on a blood sample with a negative result, immediately vaccinated for EVA, 
kept for 21  days following  vaccination  separated from other equids and regularly 
revaccinated in accordance with the recommendations of the manufacturer; or  
4. were subjected to a test for EVA on a blood sample with a negative result within 14  days 
prior to semen collection, and had been separated from other equids not of an equivalent 
EVA status for 14  days prior to blood sampling until the end of semen collection; or  
5. have been subjected to a test for EVA carried out on a blood sample with a posit ive result 
and then: either  
c) were subsequently test mated to two mares within six  months prior to semen 
collection, which were subjected to two tests for EVA with negative results on blood 
samples collected at the time of test mating and again 28  days after  the test mating; 
or 
d) were subjected to a test for EVA with a negative result, carried out on semen 
collected within six  months prior to collection of the semen to be exported; or  
e) were subjected to a test for EVA with a negative result, carried out on semen  
collected within six  months after the blood sample was collected, then immediately 
vaccinated, and regularly revaccinated; or  
6. for frozen semen, were subjected with negative results either:  
a) to a test for EVA carried out on a blood sample taken not earlier than 14  days and 
not later than 12  months after the collection of the semen for export; or  
b) to a test for EVA carried out on an aliquot of the semen collected immediately prior 
to processing or on an aliquot of semen collected within 14 to 30  days after the  first 
collection of the semen to be exported.  
Current Code recommendations for the importation of in vivo  derived equine embryos   
10  Risk Management Proposal for Equine Semen and Embryos  Ministry for Primary Industries  Veterinary Authorities  of importing countries  should require the presentation of an international 
veterinary certificate  attesting that the donor animals  showed no clinical sign of EVA on the day of 
embryo collection; and  
EITHER  
1. were kept in an establishment  where no animals  have shown any signs of EVA for the 28 days 
prior to collection; and  
a. were subjected to a test for EV A carried out on blood samples collected either once within 
21 days prior to collection with negative results, or on two occasions at least 14 days apart 
within 28 days prior to collection, which demonstrated stable or declining antibody titres; or  
b. were re gularly vaccinated according to the recommendations of the manufacturer;  
OR 
2. were isolated for the 28 days prior to collection and during this period the animals  showed no sign 
of EVA;  
AND  
3. semen used to fertilise  the oocytes complies with the requirements in the Code Article relevant to 
EVA in semen.  
5.5.3 Recommendation  
Semen  
 
Donors must meet the OIE Code recommendations for managing  EVA in equine semen . 
 
Embryo s 
 
Donors must meet the OIE Code  recommendations for managing EVA in in vivo  derived embryos . 
 
5.6 Leptospir a species  
5.6.1 Options for management of risk presented in the 2009 IRA for equine germplasm:  
Option 1  
Donor horses could be tested serologically with a variety of antigens that occur in the exporting country and not in 
New Zealand, with negative results.  
Option 2  
Donor horses could be treated with effective antibiotics within one week prior to germplasm c ollection.  
Option 3  
Diluents containing antibiotics that are effective against Leptospira  spp. could be used in the preparation of the 
semen and antibiotics could be included in the solutions used in the preparation of embryos.  
NB: this reflects the recommendations of the IETS Manual and the current IHS s for horse semen from countries 
covered by this risk analysis.  
5.6.2 Discussion  
The OIE Code  does not make any recommendations for leptospirosis. Leptospira  species  are sensitive to a 
variety of antibiotics, and treatment of animals or inclusion of antibiotics in prepared semen has traditionally been  
Ministry for Primary Industries   Risk Management Proposal for Semen and Embryos from Horses ( Equidae )  11 used to prevent dissemination of Leptospira  species  by international trade. Treatment of embryos is also likely to 
be effective.  
 
The June 2009 Review of leptospirosis measures in Import Health Standards (p.13, recommendation 2) 
recommends that semen and embryos be prepared according to the OIE Code  and International Embryo Transfer 
Society (IETS) standard.  
The OIE Manua l has a chapter on “Collection and processing of bovine, small ruminant and 
porcine semen”, but none specifically for equine semen, which includes the following:  
A mixture of antibiotics should be included with a bactericidal activity at least equivalent t o that 
of the following mixtures in each ml of frozen semen: gentamicin (250  µg), tylosin (50  µg), 
lincomycin –spectinomycin (150/300  µg); penicillin (500  IU), streptomycin (500  µg), lincomycin -
spectinomycin (150/300  µg); or amikacin (75  µg), divekacin (25  µg). 
 
From  IETS Manual  Ch 7:  
Penicillin (100 IU/mL) and str eptomycin (100 μg/mL) or gentamy cin (50 μg/mL) have been used 
most commonly to prevent bacterial replication during culture of bovine embryos.  
 
Leptospira borgpetersenii serovar hardjobovis has bee n found associated with washed oocytes 
and embryos, supplementation of the media with penicillin and streptomycin will ensure that 
embryos are not contaminated with infectious bacteria.  
 
Current IHS for equine semen  
 
An effective combination of antibiotics was added to the semen extender/diluent. The 
combination must produce an effect at least equivalent to the following:  
500 IU per ml streptomycin; or  
500 IU per ml penicillin; or  
150 µg per ml lincomycin; or  
300 µg per ml spectinomycin; or  
50 µg per ml gentamycin.  
Additionally, equivalence has been given for Ticarcillin at 1.2 mg/ml and Amikacin 0.5 mg/ml  (equals 1 .0 mg per 
ml of Timentin) . The antibiotics are listed in the MPI -Document for Approved Diagnostic Tests, Vaccines, 
Treatments and Post-Arrival Testing Laboratories for Animal Import Health Standards  (MPI-STD-TVTL ). 
5.6.3 Recommendation  
 
Semen and embryos must be prepared with antibiotics effective against Leptospira  species  as list ed in MPI -STD-
TVTL . 
 
5.7 Salmonell a species  
5.7.1 Options for management of risk presented in the 2009 IRA for equine germplasm:  
Option 1  
i. The donors were kept for the 3 months prior to collection on premises where salmonellosis has not 
occurred during that period; and  
ii. The horses were showing no clinical signs of salmonellosis on the day of collection.  
  
12  Risk Management Proposal for Equine Semen and Embryos  Ministry for Primary Industries  NB: this option reflects the current import health standards for equine semen.  
Option 2  
i. donors could be resident for at least 3 months on properties on which no cases of salmonellosis have 
been diagnosed during the previous 3 years; and  
ii. faecal samples from donors could be cultured according  to methods recommended in the OIE Manual 
of Diagnostic Tests and Vaccines (Davies 2008), three times at weekly intervals immediately before 
germplasm collection, with negative results.  
  
NB. The 3 year property freedom period and the intervals and frequency of sampling are conservative, albeit 
arbitrary.  
Option 3  
Aliquots of each semen and embryo batch to be imported could be cultured, using methods described in the OIE 
Manual of Diagnost ic Tests and Vaccines , with negative results.  
5.7.2 Discussion  
Salmonella spp.  (exotic affecting animals) are listed as exotic, unwanted organisms. There are no OIE Code  
measures recommended for Salmonella  in horses or semen or embryos. While  S. abortus equi is thought to be 
transmitted venereally, the likelihood of introducing exotic salmonellae in equine collected, processed and stored 
following OIE Code  recommendations is considered remote. There is no evidence that Salmonellosis has been 
transmitted via seme n or embryos collected and processed in such a manner.   
Option 1 is consistent with the current requirements for importing horses:   
The horses were showing no clinical signs of equine Salmonellosis at the final inspection prior 
to departure and were kept for at least the past 90 days on premises where no case of equine 
Salmonellosis ( S. abortus equi ) has been reported during that time.  
Option 2 is consistent with the current OIE Code  in applying no measures.  
5.7.3 Recommendation  
 
It is recommended no measures r egarding equine salmonellosis are applied to either semen or embryos.  
 
5.8 Taylorella species (C ontagious equine metritis , CEM ) 
5.8.1 Options for management of risk presented in the 2009 IRA for equine germplasm:  
Option 1  
Donors of germplasm could be required to be resident for at least 2 months on a property that is not considered 
to be an infected establishment according to the Code  definition.  
Option 2  
a) Donors of germplasm could be required to be resident for at least 2 months on a property that is not considered 
to be an infected establishment according to the Code  definition; and  
b) Swabs from the donors could be tested, with negative results, by culturing or PCR on three occasions. Once 
within 7 days prior to germplasm c ollection and twice at weekly intervals during the 21 days after germplasm 
collection. The swabs could be collected from the prepuce, urethral sinus and fossa glandis (including the 
diverticulum) of stallions and from the mucosal surfaces of the urethra, c litoral sinuses and clitoral fossa in the 
case of mares .  
Option 3    
Ministry for Primary Industries   Risk Management Proposal for Semen and Embryos from Horses ( Equidae )  13 Germplasm could be sourced from any donors provided semen diluents and solutions used for preparation of 
embryos contain antibiotics that are effective against Taylorella spp.  
Option 4  
Germplasm could be imported from countries in which CEM occurs, but is notifiable, provided that donors have 
been resident for at least 3 months on a property on which no case of CEM has been found for at least 3 years.  
Option 5  
Germplasm imports could be  restricted to donors from countries that are considered free from CEM on the basis 
that the disease is notifiable and has not occurred in the previous 3 years.  
5.8.2 Discussion  
Taylorella  equigenitalis  is an unwanted, notifiable  organism.  The OIE Manual  ident ifies this as the causative 
agent of contagious equine metritis  (CEM) .  As the organism is known to be transmitted venereally, measures are 
justified.  The Manual  also notes that T. asinigenitalis  has been  identified in male donkeys , mares and stallions, 
and “would appear to be passed to other donkeys and horses during mating”. It has not been associated with 
naturally occurring disease. A PCR test is required to distinguish between these two species.   
A recent review of CEM measure s for the revision of the Import Health Standard for Horses (1 February 
2013) resulted in a recommendation to align with the current OIE  Code  and Manual .   
The Manual  recommends culture as the method of identification, which does not distinguish between th e two 
species of Taylorella . MPI has approved quantitative polymerase chain reaction (qPCR) as a testing method in 
horses. This recommendation will be adopted in the semen and embryos standards as well.  
 
The 2012 OIE Code  has the following measures for live horses:  
Recommendations for the importation of stallions and mares considered free from CEM (for 
countries where an official control organisation is present)  
Veterinary Authorities  of importing countr ies should require the presentation of an 
international veterinary certificate  attesting that the animals : 
1. showed no clinical sign of CEM on the day of shipment;  
2. have had no contact with CEM:  
a. directly, through coitus with an infected animal ; or 
b. indirectly, by passing through an infected establishment ; 
3. were subjected to the laboratory  test for CEM with negative results during the 30  days 
prior to shipment.  
Article 12.2.3.  
Recommendations for the importation of stallions and mare s which have previously shown 
signs of CEM or which have been in contact with CEM (for countries where an official control 
organisation is present)  
Veterinary Authorities  of importing countries  should require the presentation of an 
international veterinary certificate  attesting that the animals  which have been in direct contact 
through coitus with an infected animal , or indirect contact by passing through an infected 
establishment : 
1. have been recognised as not being contagious through laboratory tests for CEM;  
2. have been protected against any possibility of contagion since the beginning of the tests.  
 
  
14  Risk Management Proposal for Equine Semen and Embryos  Ministry for Primary Industries  May 2014 MPI Import Health Standard for Horses requires   
 
Either  
a. were from a CEM -free country, approved by MPI; or  
b. were considered free from CEM; an official control programme for CEM or MPI -approved 
equivalent is established in the country of export; and horses have met the 
recommendations as described in the OIE Code  and OIE Manual (or pre -mating testing 
followed rec ommendations in the OIE Manual but was done in the 60 days prior to 
mating or artificial insemination); or  
c. have been known to be infected with CEM and were subject to an effective method of 
treatment and testing approved by MPI.  
 
To align with the OIE Code where possible, the measures imposed on semen and embryos 
from horses should reflect the recommendations for importing live horses as described in the 
Code . The Code  defines horses as infected if they have ever been tested positive for CEM 
or been in di rect or indirect contact with infected horses or establishments. It also 
recommends an establishment can be declared free two months after the confirmation of the 
last case and after thorough cleaning and disinfection of the premises. The Manual  briefly 
describes characteristics of a control programme, based on import and export testing and 
testing for breeding populations.  
5.8.3 Recommendation  
 
Donors must meet measures reflecting the OIE Code recommendations for managing CEM in horses .