**7. Documentation**

Over the years of existence of the KETRI Cryo‐bank, the cryopreserved materials were documented manually on specially designed record sheets known as Kalamazoo and later electronically. Whichever method is used, the information in the records should include, but not limited to the following, for the ease of retrieval:


#### **7.1. Electronic database**

**•** Tissue culture

16 Cryopreservation in Eukaryotes

**•** Molecular biology studies:

do not grow in rodents.

**7. Documentation**

**•** Date of isolation

**•** Isolating scientist

morphology)

**•** Locality of isolation

**◦** DNA and RNA extraction and analysis

**◦** Genetics and functional genomic studies

not limited to the following, for the ease of retrieval:

**•** The host of isolation, age and sex

ance of trypanosomes in the mouse

*Trypanosoma evansi* does not transform into procyclics in culture.

**•** *In vivo* studies using various animals models to characterize virulent phenotypes and drug resistance. It is important to note that some trypanosome species such as *Trypanosoma vivax*

Over the years of existence of the KETRI Cryo‐bank, the cryopreserved materials were documented manually on specially designed record sheets known as Kalamazoo and later electronically. Whichever method is used, the information in the records should include, but

**•** Suspected species of trypanosome (based on host of isolation and trypanosome

**•** If by propagation in laboratory rodents, the species of rodent used

sacrifice of the animal to harvest trypanosomes for cryopreservation

**•** Work carried out and publications resulting from the use of stabilates

and duration of infection must be stated at each passage.

**•** Physical location of the stabilate in the cryo‐bank

**•** Method of cryopreservation whether direct or after propagation in laboratory animals

**•** The prepatent period, i.e., period between inoculation of the whole blood and first appear‐

**•** Duration of infection before cryopreservation, i.e., the period between inoculation and the

**•** In the event where several passages have been made, the passage numbers must be indicated as well as the species of rodents used at every passage. In addition, the pre‐patent period The database was developed using Microsoft Access 2000 (Microsoft, USA) relational database. Hosts of isolation and countries are coded following the International Organization for Standardization (ISO) protocol [26] (Lumsden, 1978). Primary isolates are, for example, designated MHOM/KE/85/KETRI 128, where M represents mammal; HOM represents human; KE indicates the country of isolation, in this case Kenya; 85 is the year of isolation (1985) while KETRI 128 shows code or reference number of the stabilate. With regard to trypanosome derived from the previous stabilate usually referred to as derivatives, the number of the derivative is shown in brackets. For example, MHOM/KE/85/KETRI 128 [KETRI 300] is a derivative of MHOM/KE/85/KETRI 128 as described by Lumsden [25].

#### **7.2. Trypanosomes isolated, documented, and cryopreserved at KETRI Cryo‐bank**

The number of trypanosomes isolated and cryopreserved at the KETRI Trypanosome Bank are shown in **Figure 8**. **Tables 2** and **3** show the trypanosome species, year, and country of isolation.

**Figure 8.** Number of primary trypanosome stabilates collected, preserved, and stored at Kenya [1].



Adapted from Murilla et al. [1].

*Tbb*, *Trypanosoma bruceibrucei*; *Tbr*, *Trypanosoma bruceirhodesiense*; *Tbg*, *Trypanosoma bruceigambiense*; UN, unclassified; NDA, no data available.

**Table 2.** Primary trypanosome isolates collected from various countries and stored at the Kenya Trypanosomiasis Research Institute Cryo‐bank


Adapted from Murilla et al. [1].

**Table 3.** Animal hosts from which various trypanosomes were isolated and stored at Kenya Trypanosomiasis Research Institute Cryo‐bank

#### **8. Issuance of stabilates**

Trypanosomes are stored and maintained permanently in liquid nitrogen for use by scientists in the following areas:


Stabilates are issued to National and International Research Organizations and Institutions of higher learning following the laid down institutional guidelines. The following documents are necessary for issue of materials to be effected:

#### **Issuance of trypanosome stabilates**

#### **Procedure**

**Species of trypanosomes: number and period of isolation**

Mozambique No – – 2 – – – – – – – – –

Nigeria No – – – – – 4 – – – – – –

Zambia No – – – – 2 – – – – – – –

NDA No 2 21 8 – 17 5 1 – – – 3 2

Total 104 510 416 48 243 240 92 3 2 8 35 45

**Table 2.** Primary trypanosome isolates collected from various countries and stored at the Kenya Trypanosomiasis

Cattle 85 247 0 119 137 0 2 0 10 21 621 Goat 0 6 0 6 6 0 0 0 0 0 1 19 Sheep 1 8 1 24 3 0 0 0 0 0 7 44 Pig 1 5 0 0 0 0 0 0 0 0 0 6 Camel 0 3 0 1 2 92 0 0 0 0 0 98 Donkey 0 1 0 1 0 0 0 0 0 3 0 5 Cat 0 0 0 0 0 0 0 0 0 0 2 2 Dog 3 6 0 1 0 0 0 0 0 1 0 11 Wildlife 2 40 3 12 0 0 0 0 0 0 5 62 Lizard 0 0 0 0 0 0 0 0 0 3 0 3 Rat 0 0 0 0 0 0 0 0 8 0 0 8 HNI 1 7 4 2 2 0 0 0 0 2 0 18 Total 93 323 8 166 150 92 2 0 8 19 36 897

**Table 3.** Animal hosts from which various trypanosomes were isolated and stored at Kenya Trypanosomiasis Research

Trypanosomes are stored and maintained permanently in liquid nitrogen for use by scientists

*Tbb*, *Trypanosoma bruceibrucei*; *Tbr*, *Trypanosoma bruceirhodesiense*; *Tbg*, *Trypanosoma bruceigambiense*; UN, unclassified; NDA,

*congolense*

*T. vivax T. evansi T. simiae T. theileri*

1973

Year – – – – 1981 – – – – – – –

Year 1961 – – – 1962–1985 1961 – – – – – 2

*Tbb Tb subgroup Tbr T. congolense T. vivax T. evansi T. theileri T. simiae T. lewesi UC Mixed Total*

– – – – – – – – –

*T. lewesi*

– – – – – –

**UC Mixed**

*Tbr Tbg T.*

1983

Year – – – – – 1970–

**Country Isolate/**

18 Cryopreservation in Eukaryotes

**Year**

Adapted from Murilla et al. [1].

Research Institute Cryo‐bank

Adapted from Murilla et al. [1].

**8. Issuance of stabilates**

**2.** Molecular characterization

**1.** The pathogenicity/virulence studies

in the following areas:

Institute Cryo‐bank

no data available.

*Tbb Tb*

Year – – 1980,

*subgroup*


It is important that the receiving scientist be committed to avail the scientific information resulting from the use of these stabilates. This is necessary for updating the trypanosome bank database and for future reference.
